Homotypic RNA clustering accompanies a liquid-to-solid transition inside the core of multi-component biomolecular condensates
Data files
Jul 16, 2025 version files 2.71 GB
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Dryad_source_data_for_extended_data.zip
29.01 MB
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Dryad_source_data_for_main_text.zip
92.27 MB
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Dryad_source_data_for_supplementary_information.zip
2.59 GB
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README.md
31.49 KB
Abstract
RNA-driven condensation plays a central role in organizing and regulating ribonucleoprotein granules within cells. Disruptions to this process, such as the aberrant aggregation of repeat-expanded RNA, are associated with numerous neurological disorders. Here, we report that irreversible RNA aggregation is facilitated within multicomponent protein–nucleic acid condensates. Using time-lapse confocal fluorescence microscopy, we tracked the growth of RNA clusters as a function of condensate age and found that RNA aggregation is driven by RNA percolation, proceeding via a transition from intramolecular to intermolecular RNA–RNA interactions. For GC-rich repeat RNAs, the size of RNA clusters increases, and the timescale of cluster formation decreases with the number of repeat units. Utilizing nanorheology, optical tweezer-based droplet fusion assays, and high-resolution confocal fluorescence imaging, we uncovered that RNA clustering drives a liquid-to-solid phase transition in the condensate core, leading to the emergence of multiphasic condensate structures. Multivalent RNA-binding cofactors, such as antisense oligonucleotides and RNA-binding proteins, that compete with homotypic RNA–RNA interactions can raise the activation energy barrier for RNA clustering, thereby acting as inhibitors of intra-condensate RNA aggregation. The insights gained from this study provide a complementary perspective on the role of multicomponent biomolecular condensates in regulating aberrant RNA self-assembly in living cells.
This readme file was generated on 2025-07-15 by Tharun Selvam Mahendran
DESCRIPTION:
The dataset provided here contains imaging data (figures/videos) corresponding to the main text, extended data, and supplementary information concerning the referenced publication. Methodological details and additional acquisition parameters can be found in the relevant methods section of the publication.
The images are organized into folders, broadly categorized into main text figures (Dryad_source_data_for_main_text.zip), extended data figures (Dryad_source_data_for_extended_data.zip), and supplementary information figures/videos (Dryad_source_data_for_supplementary_information.zip). The main folders include subfolders specified for individual image panels if the concerned figure includes multiple panels of data. These subfolders are named following the corresponding panel letter, e.g., a, b, c, d, etc.
Further, the series of images constituting a figure are categorized according to experimental parameters if multiple image data exists in a single figure panel: timepoint of imaging (mentioned in hours, minutes or seconds), fluorophore being imaged (e.g., SYTO13, Alexa594 or A594, Cy5, FAM, ThT), and sample composition (e.g., RGG peptide, dT40 ssDNA, TERRA RNA, CAG31 repeat RNA), etc. A description is provided along with each set of files to provide clarification on technical terms used. Additionally, a direct correspondence exists between the organized data in the repository and the data shown in the published article. Therefore, details of the sample and additional information about the sample conditions, if applicable, can be found in the publication, specifically, the relevant figure legend and/or method section.
GENERAL INFORMATION:
Title of Dataset: Homotypic RNA clustering accompanies a liquid-to-solid transition inside the core of multi-component biomolecular condensates
Author/Principal Investigator Information
Name: Priya R. Banerjee
ORCID: https://orcid.org/0000-0002-6169-1461
Institution: The State University of New York at Buffalo
Address: 229 Fronczak Hall
Email: prbanerj@buffalo.edu
Author
Name: Tharun Selvam Mahendran
ORCID: https://orcid.org/0000-0002-7536-1236
Institution: The State University of New York at Buffalo
Address: 206 Fronczak Hall
Email: tharunse@buffalo.edu
Author/Alternate Contact Information
Name: Anurag Singh
ORCID: https://orcid.org/0000-0002-8132-903X
Institution: The State University of New York at Buffalo
Address:
Email: singh26@buffalo.edu
Author/Alternate Contact Information
Name: Gable M Wadsworth
ORCID: https://orcid.org/0000-0002-0417-9613
Institution: The State University of New York at Buffalo
Address:
Email: gablewad@buffalo.edu
Date of data collection: between 2023-02-01 and 2025-01-20
Geographic location of data collection: Buffalo, New York, United States of America
Information about funding sources that supported the collection of the data: This work was supported by the US National Institutes of Health through grants R35 GM138186 (P.R.B)
and the St. Jude Children’s Research Collaborative on the Biology and Biophysics of RNP Granules (P.R.B.).
SHARING/ACCESS INFORMATION
Licenses/restrictions placed on the data: CC Zero
Links to publications that cite or use the data: https://www.nature.com/articles/s41557-025-01847-3
Links to other publicly accessible locations of the data:
Links/relationships to ancillary data sets: https://github.com/BanerjeeLab-
repertoire/Biomolecular-Condensates-Can-Enhance-Pathological-RNA-Clustering
Was data derived from another source? NO
If yes, list source(s):
Recommended citation for this dataset:
Mahendran, T.S., Wadsworth, G.M., Singh, A. et al. Homotypic RNA clustering accompanies a liquid-to-solid transition inside the core of multi-component biomolecular condensates. Nat. Chem. (2025). https://doi.org/10.1038/s41557-025-01847-3
Usage notes: File types are either in .tif or .png formats. Either file type can be accessed via Fiji/ImageJ, a freely available software.
DATA & FILE OVERVIEW
File List:
>Source data for the main text (contains all the microscopy data shown in the main text)
Dryad_source_data_for_main_text.zip
Figure 1
- c
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0 hr.tif
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24 hr.tif
Description: These image files correspond to different time points. hr indicates hours.
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- d
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Merge.tif
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Cy5 dT40.tif
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A594 RGG.tif
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SYTO-13.tif
Description: These image files correspond to different fluorescence channels, e.g., Cy5 dT40 refers to fluorescence channel corresponding to Cy5 dT40 excitation/emission.
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- e
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1.5 hr.tif
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2 hr.tif
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5 hr.tif
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5.25 hr.tif
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5.5 hr.tif
Description: These image files correspond to different time points. hr indicates hours.
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- f
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Merge.tif
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A594-RGG.tif
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SYTO-13.tif
Description: These image files correspond to different fluorescence channels, e.g., A594-RGG refers to fluorescence channel corresponding to A594-RGG excitation/emission.
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- h
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Merge.tif
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Cy5-dT40.tif
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SYTO-13.tif
Description: These image files correspond to different fluorescence channels, e.g., Cy5-dT40 refers to fluorescence channel corresponding to Cy5 dT40 excitation/emission.
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- j
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z stacks.tif
Description: Individual z slices in the form of an image stack.
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Figure 2
- b
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20C.tif
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90C.tif
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Cooled to 20C.tif
Description: 'C' here refers to degrees Celsius, e.g., 20C refers to 20 degrees Celsius.
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- d
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1st from left.tif
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2nd from left.tif
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3rd from left.tif
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4th from left.tif
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5th from left.tif
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6th from left.tif
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7th from left.tif
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8th from left.tif
Description: The naming order described here is in reference to a series of images, corresponding to imaging data from measurements involving temperature ramps. The '1st from left.tif' is the start of the order and '8th from left.tif' is the end.
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- e
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20C.tif
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40C.tif
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Cooled to 20C.tif
Description: 'C' here refers to degrees Celsius, e.g., 20C refers to 20 degrees Celsius.
-
- g
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0 hr SYTO-13.tif
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0 hr A594-RGG.tif
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0 hr Merge.tif
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24 hr SYTO-13.tif
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24 hr A594-RGG.tif
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24 hr Merge.tif
Description: These image files correspond to distinct time points and fluorescence channels. For example, 0 hr SYTO-13 refers to the 0-hour time point
with SYT0-13 fluorescence channel.
-
- h
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TERRAx10 0hr.tif
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TERRAx10 24hr.tif
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mut-TERRAx10 0hr.tif
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mut-TERRAx10 24hr.tif
Description: These image files correspond to the RNA used in sample preparation and the time point. For example, TERRAx10 0hr refers to the RNA, TERRAx10, and the time point of 0 hours; 'x10' in TERRAx10 refers to number of RNA repeat units.
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Figure 3
- a
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TERRAx1 0hr.tif
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TERRAx1 2hr.tif
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TERRAx1 4hr.tif
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TERRAx1 8hr.tif
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TERRAx1 18hr.tif
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TERRAx4 0hr.tif
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TERRAx4 2hr.tif
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TERRAx4 4hr.tif
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TERRAx4 8hr.tif
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TERRAx4 18hr.tif
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TERRAx6 0hr.tif
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TERRAx6 2hr.tif
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TERRAx6 4hr.tif
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TERRAx6 8hr.tif
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TERRAx6 18hr.tif
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TERRAx10 0hr.tif
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TERRAx10 2hr.tif
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TERRAx10 4hr.tif
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TERRAx10 8hr.tif
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TERRAx10 18hr.tif
Description: These image files correspond to the RNA used in sample preparation and the time point. For example, TERRAx10 0hr refers to the RNA, TERRAx10, and the time point of 0 hours; 'x10' in TERRAx10 refers to the number of RNA repeat units.
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- b
- Image 0 hr
- spatial autocorrelation 0 hr
- Image 18 hr
- spatial autocorrelation 18 hr
- Description: These files are categorized either as the image files or the corresponding spatial autocorrelation plots with the time point indicated wherein hr means hours.
Figure 4
- b
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CAGx20 SYTO-13 0hr.tif
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CAGx20 A594 RGG 0hr.tif
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CAGx20 Merge 0hr.tif
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CAGx20 SYTO-13 24hr.tif
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CAGx20 A594 RGG 24hr.tif
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CAGx20 Merge 24hr.tif
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CAGx31 SYTO-13 0hr.tif
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CAGx31 A594 RGG 0hr.tif
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CAGx31 Merge 0hr.tif
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CAGx31 SYTO-13 24hr.tif
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CAGx31 A594 RGG 24hr.tif
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CAGx31 Merge 24hr.tif
Description: These image files correspond to the RNA used in sample preparation, the fluorescence channel, and the time point. For example, CAGx31 SYTO-13 0hr refers to the RNA, CAGx31; the fluorescence channel, SYTO-13; the time point of 0 hours; 'x31' in CAGx31 refers to the number of RNA repeat units.
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- d
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CUGx31 SYTO-13 0hr.tif
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CUGx31 A594 RGG 0hr.tif
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CUGx31 Merge 0hr.tif
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CUGx31 SYTO-13 24hr.tif
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CUGx31 A594 RGG 24hr.tif
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CUGx31 Merge 24hr.tif
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CUGx47 SYTO-13 0hr.tif
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CUGx47 A594 RGG 0hr.tif
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CUGx47 Merge 0hr.tif
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CUGx47 SYTO-13 24hr.tif
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CUGx47 A594 RGG 24hr.tif
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CUGx47 Merge 24hr.tif
Description: These image files correspond to the RNA used in sample preparation, the fluorescence channel, and the time point. For example, CAGx31 SYTO-13 0hr refers to the RNA, CAGx31; the fluorescence channel, SYTO-13; the time point of 0 hours; 'x31' in CAGx31 refers to the number of RNA repeat units.
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Figure 5
- a
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20 min before fusion.tif
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20 min after fusion.tif
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150 min before fusion.tif
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150 min after fusion.tif
Description: Here, the files are organized in terms of time point and the condition of 'after fusion' and 'before fusion'. For example, 20 min before fusion refers to 20 minute time point, and the image collected before fusion event
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- b
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Before.tif
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6 s.tif
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8 s.tif
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10 s.tif
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16 s.tif
Description: Here, the files are organized according to time point wherein s refers to seconds. 'Before' refers to the initial time point or 0 s data. For example, 8 s refers to 8 seconds.
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Figure 6
- b
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FAM TERRAx4 0hr.tif
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A594 RGG 0hr.tif
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Merge 0 hr.tif
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FAM TERRAx4 24hr.tif
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A594 RGG 24hr.tif
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Merge 24 hr.tif
Description: These image files correspond to the fluorescence channel, and the time point. For example, FAM TERRAx4 0hr refers to the fluorescence channel, FAM TERRAx4; the time point of 0 hours; 'x4' in FAM TERRAx4 refers to the number of RNA repeat units.
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- d
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before.tif
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1 hr.tif
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2.25 hr.tif
Description: Here, the files are organized according to time point wherein hr refers to hours. 'Before' refers to the initial time point or 0 hr data. For example, 1 hr refers to 1 hour.
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- f
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FAM TERRAx4 0hr.tif
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A594 RGG 0hr.tif
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Merge 0 hr.tif
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FAM TERRAx4 24hr.tif
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A594 RGG 24hr.tif
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Merge 24 hr.tif
Description: These image files correspond to the fluorescence channel, and the time point. For example, FAM TERRAx4 0hr refers to the fluorescence channel, FAM TERRAx4; the time point of 0 hours; 'x4' in FAM TERRAx4 refers to the number of RNA repeat units.
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- i
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SYTO13 0hr.tif
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A594 RGG 0hr.tif
-
Merge 0 hr.tif
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SYTO13 24hr.tif
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A594 RGG 24hr.tif
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Merge 24 hr.tif
Description: These image files correspond to the fluorescence channel, and the time point. For example, SYTO13 0hr refers to the fluorescence channel, SYTO13; the time point of 0 hours.
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- j
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SYTO13 0hr.tif
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A594 RGG 0hr.tif
-
Merge 0 hr.tif
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SYTO13 24hr.tif
-
A594 RGG 24hr.tif
-
Merge 24 hr.tif
Description: These image files correspond to the fluorescence channel, and the time point. For example, SYTO13 0hr refers to the fluorescence channel, SYTO13; the time point of 0 hours.
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>Source data for supplementary text (contains all the microscopy data shown in the supplementary text including videos)
Dryad_source_data_for_supplementary_information.zip
>Supplementary Figures
Figure 1
-
24 hr.tif
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0 hr.tif
Description: The files correspond to images collected at different time points. For example, 0 hr refers to 0 hours.
Figure 2
-
24 hr.tif
-
0 hr.tif
Description: The files correspond to images collected at different time points. For example, 0 hr refers to 0 hours.
Figure 3
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SYTO-13 0hr.tif
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A594-RGG 0hr.tif
-
Merge 0 hr.tif
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SYTO-13 24hr.tif
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A594-RGG 24hr.tif
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Merge 24 hr.tif
Description: These image files correspond to the fluorescence channel, and the time point. For example, SYTO-13 0hr refers to the fluorescence channel, SYTO-13; the time point of 0 hours.
Figure 4
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Image sequence of timelapse.tif
Description: A sequence of fluorescence images constituting a timelapse from beginning to end.
Figure 5
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5%.tif
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10%.tif
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20%.tif
Description: Here the files are organized in terms of the crowder percentage.
Figure 6
- a
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0 ugml.tif
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8 ugml.tif
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78 ugml.tif
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196 ugml.tif
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314 ugml.tif
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444 ugml.tif
Description: Here, the files are organized in terms of the RNA concentration expressed as ugml, which means micrograms/milliliters.
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- e
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0 hr.tif
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24 hr.tif
Description: Here, the files are organized in terms of time points.
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Figure 8
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0 hr SYTO-13.tif
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0 hr Cy5-dT40.tif
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0 hr Merge.tif
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24 hr SYTO-13.tif
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24 hr Cy5-dT40.tif
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24 hr Merge.tif
Description: These files are organized in terms of time point and fluorescence channel. For example, 0 hr SYTO-13 refers to 0 hour timepoint and SYTO-13 fluorescence channel.
Figure 10
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RGG dT40 condensates 0 hr.tif
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RGG dT40 condensates 24 hr.tif
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RGG dT40 polyU condensates 0 hr.tif
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RGG dT40 polyU condensates 24 hr.tif
Description: These image files are organized according to the condensate sample, and the time point. For example, RGG dT40 condensates 0 hr refers to the RGG dT40 condensates and the time point of 0 hours.
Figure 11
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NaCl treated condensates SYTO-13 channel 0 hr.tif
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NaCl treated condensates Cy5-dT40 channel 0 hr.tif
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NaCl treated condensates Merge 0 hr.tif
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NaCl treated condensates SYTO-13 channel 2.5 hr.tif
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NaCl treated condensates Cy5-dT40 channel 2.5 hr.tif
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NaCl treated condensates Merge 2.5 hr.tif
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NaCl treated condensates SYTO-13 channel 24 hr.tif
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NaCl treated condensates Cy5-dT40 channel 24 hr.tif
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NaCl treated condensates Merge 24 hr.tif
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KCl treated condensates SYTO-13 channel 0 hr.tif
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KCl treated condensates Cy5-dT40 channel 0 hr.tif
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KCl treated condensates Merge 0 hr.tif
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KCl treated condensates SYTO-13 channel 2.5 hr.tif
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KCl treated condensates Cy5-dT40 channel 2.5 hr.tif
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KCl treated condensates Merge 2.5 hr.tif
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KCl treated condensates SYTO-13 channel 24 hr.tif
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KCl treated condensates Cy5-dT40 channel 24 hr.tif
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KCl treated condensates Merge 24 hr.tif
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LiCl treated condensates SYTO-13 channel 0 hr.tif
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LiCl treated condensates Cy5-dT40 channel 0 hr.tif
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LiCl treated condensates Merge 0 hr.tif
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LiCl treated condensates SYTO-13 channel 2.5 hr.tif
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LiCl treated condensates Cy5-dT40 channel 2.5 hr.tif
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LiCl treated condensates Merge 2.5 hr.tif
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LiCl treated condensates SYTO-13 channel 24 hr.tif
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LiCl treated condensates Cy5-dT40 channel 24 hr.tif
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LiCl treated condensates Merge 24 hr.tif
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NH4OAc treated condensates Cy5-dT40 channel 0 hr.tif
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NH4OAc treated condensates Merge 0 hr.tif
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NH4OAc treated condensates SYTO-13 channel 2.5 hr.tif
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NH4OAc treated condensates Cy5-dT40 channel 2.5 hr.tif
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NH4OAc treated condensates Merge 2.5 hr.tif
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NH4OAc treated condensates SYTO-13 channel 24 hr.tif
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NH4OAc treated condensates Cy5-dT40 channel 24 hr.tif
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NH4OAc treated condensates Merge 24 hr.tif
Description: The files are organized in terms of chemical treatment, fluorescence channel, and time points. For example, LiCl treated condensates Merge 24 hr refers to a sample with LiCl treatment, the Merge channel, and 24 hours time point.
Figure 12
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panel A
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panel B
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panel C
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panel D
Description: Each file represents an image or plot corresponding to a particular panel: A, B, C, and D.
Figure 13
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panel a Image final xy.png
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panel a Image final xz.png
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panel a Image final yz.png
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panel b Confocal image - 100nm.png
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panel b Heat map - 100nm.png
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panel b spatial autocorrelation - 100nm.png
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panel c Confocal image - 200nm.png
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panel c Heat map - 200nm.png
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panel c spatial autocorrelation - 200nm.png
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panel d Confocal image - 1um.png
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panel d Heat map - 1um.png
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panel d spatial autocorrelation - 1um.png
Description: Here the files are organized in terms of the panel letter and whether the data corresponds to either an image, heat map, or spatial autocorrelation, as well as the size of the bead, such as 100 nm, 200 nm, and 1 um; nm means nanometers and um means micrometers.
Figure 14
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panel a FAM-TERRA 0 hr.tif
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panel a A594-RGG 0 hr.tif
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panel a FAM-TERRA 4 hr.tif
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panel a A594-RGG 4 hr.tif
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panel a FAM-TERRA 8 hr.tif
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panel a A594-RGG 8 hr.tif
Description: Here the files are organized in terms of the panel letter, fluorescence channel and the time point. For example, panel a FAM-TERRA 0 hr corresponds to data in panel a, FAM-TERRA channel and 0 hour time point.
Figure 15
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Panel a 0.1mg/ml CAGx31 SYTO-13.tif
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Panel a 0.1mg/ml CAGx31 A594-RGG.tif
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Panel a 0.1mg/ml CAGx31 Merge.tif
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Panel a 0.5mg/ml CAGx31 SYTO-13.tif
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Panel a 0.5mg/ml CAGx31 A594-RGG.tif
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Panel a 0.5mg/ml CAGx31 Merge.tif
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Panel a 1mg/ml CAGx31 SYTO-13.tif
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Panel a 1mg/ml CAGx31 A594-RGG.tif
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Panel a 1mg/ml CAGx31 Merge.tif
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Panel b 0.1mg/ml CAGx31 SYTO-13.tif
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Panel b 0.1mg/ml CAGx31 Cy5-dT40.tif
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Panel b 0.1mg/ml CAGx31 Merge.tif
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Panel b 0.5mg/ml CAGx31 SYTO-13.tif
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Panel b 0.5mg/ml CAGx31 Cy5-dT40.tif
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Panel b 0.5mg/ml CAGx31 Merge.tif
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Panel b 1mg/ml CAGx31 SYTO-13.tif
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Panel b 1mg/ml CAGx31 Cy5-dT40.tif
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Panel b 1mg/ml CAGx31 Merge.tif
Description: Here the files are organized in terms of the panel letter, RNA concentration, and fluorescence channel. For example, Panel b 0.5mg/ml CAGx31 SYTO-13 corresponds to data in panel b, 0.5 mg/ml (corresponds to milligrams/milliliter) of CAGx31 RNA (x31 refers to 31 RNA repeats) and SYTO-13 fluorescence channel.
Figure 16
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0 hr SYTO-13.tif
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0 hr Cy5-dT40.tif
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0 hr Merge.tif
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24 hr SYTO-13.tif
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24 hr Cy5-dT40.tif
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24 hr Merge.tif
Description: Here the files are organized in terms of the time point and fluorescence channel. For example, 0 hr SYTO-13 refers to 0 hour time point and SYTO-13 fluorescence channel.
Figure 17
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CAGx20.tif
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CAGx31.tif
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GGGGCCx5.tif
Description: Here the files are organized according to the RNA used in the sample (CAG or GGGGCC). For example, CAGx20 means CAG RNA and here, x20 refers to the number of RNA repeat units, which is 20 in this example.
Figure 18
- b
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Before.tif
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2 s.tif
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4 s.tif
Description: Here the files are organized according to time point. 'Before' refers to the initial time point, and s refers to seconds.
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- c
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Before.tif
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2 s.tif
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4 s.tif
Description: Here the files are organized according to time point. 'Before' refers to the initial time point, and s refers to seconds.
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Figure 19
- panel a
Figure 23
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SYTO-13 0 hr.tif
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Cy5-dT40 0 hr.tif
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Merge 0 hr.tif
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SYTO-13 24 hr.tif
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Cy5-dT40 24 hr.tif
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Merge 24 hr.tif
Description: Here the files are organized in terms of the time point and fluorescence channel. For example, SYTO-13 0 hr refers to SYTO-13 fluorescence channel and 0 hour time point.
Figure 24
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0 hr FAM TERRA.tif
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0 hr Cy5 dT40.tif
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0 hr composite.tif
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24 hr FAM TERRA.tif
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24 hr Cy5 dT40.tif
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24 hr composite.tif
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48 hr FAM TERRA.tif
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48 hr Cy5 dT40.tif
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48 hr composite.tif
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72 hr FAM TERRA.tif
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72 hr Cy5 dT40.tif
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72 hr composite.tif
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96 hr FAM TERRA.tif
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96 hr Cy5 dT40.tif
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96 hr composite.tif
Description: Here the files are organized in terms of time point (hr means hours) and fluorescence channel (FAM TERRA, Cy5 dT40, composite).
Figure 26
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before.tif
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3 hr.tif
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9 hr.tif
Description: Here the files are organized in terms of time points. Before refers to the initial timepoint and hr refers to hours.
Figure 27
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A488 G3BP1.tif
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Cy5 dT40.tif
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Composite.tif
Description: Here the files are organized in terms of fluorescence channel, A488 G3BP1, Cy5 dT40, or Composite.
Figure 28
- panel a
- 0 uM
- 2.5 uM
- 5 uM
- 10 uM
- 15 uM
- 23 uM
Figure 29
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0 hr SYTO-13.tif
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0 hr Cy5-dT40.tif
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0 hr Merge.tif
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1 hr SYTO-13.tif
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1 hr Cy5-dT40.tif
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1 hr Merge.tif
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2 hr SYTO-13.tif
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2 hr Cy5-dT40.tif
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2 hr Merge.tif
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4 hr SYTO-13.tif
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4 hr Cy5-dT40.tif
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4 hr Merge.tif
Description: Here the files are organized in terms of time point (hr means hours) and fluorescence channel (SYTO-13, Cy5-dT40, Merge).
Figure 30
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TERRAx10 FAM-TERRA 0 hr.tif
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TERRAx10 Cy5-dT40 0 hr.tif
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TERRAx10 Merge 0 hr.tif
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TERRAx10 FAM-TERRA 24 hr.tif
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TERRAx10 Cy5-dT40 24 hr.tif
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TERRAx10 Merge 24 hr.tif
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CAGx31 FAM-TERRA 0 hr.tif
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CAGx31 Cy5-dT40 0 hr.tif
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CAGx31 Merge 0 hr.tif
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CAGx31 FAM-TERRA 24 hr.tif
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CAGx31 Cy5-dT40 24 hr.tif
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CAGx31 Merge 24 hr.tif
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CUGx47 FAM-TERRA 0 hr.tif
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CUGx47 Cy5-dT40 0 hr.tif
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CUGx47 Merge 0 hr.tif
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CUGx47 FAM-TERRA 24 hr.tif
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CUGx47 Cy5-dT40 24 hr.tif
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CUGx47 Merge 24 hr.tif
Description: Here the files are organized in terms of RNA in sample, fluorescence channel and time point. For example, CUGx47 FAM-TERRA 0 hr refers to CUGx47 RNA (x47 refers to number of repeat units), FAM-TERRA fluorescence channel, and the time point, 0 hours.
Figure 32
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Before addition.tif
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Post addition 4 hr.tif
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Post addition 24 hr.tif
Description: Here the files are organized in terms of before and after the addition of the reagent, imaged at 4 hours and 24 hours. hr means hours.
Figure 34
- panel a
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0 hr.tif
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24 hr.tif
Description: Here the files are organized in terms of time points. hr means hours.
-
- panel b
-
0 hr.tif
-
24 hr.tif
Description: Here the files are organized in terms of time points. hr means hours.
-
Figure 35
- a
- b
- c
>Supplementary Videos
Video 1
-
Video
Description: A series of images compiled as an image stack
Video 2
-
Video.tif
Description: A series of images compiled as an image stack
Video 3
-
Video.tif
Description: A series of images compiled as an image stack
Video 4
-
Video
Description: A series of images compiled as an image stack
Video 5
-
Video.tif
Description: A series of images compiled as an image stack
Video 6
-
video
Description: A series of images compiled as an image stack
Video 7
-
video.tif
Description: A series of images compiled as an image stack
Video 8
-
Timelapse microscopy
Description: A series of images compiled as an image stack
Video 9
-
timelapse movie
Description: A series of images compiled as an image stack
Video 10
-
timelapse movie
Description: A series of images compiled as an image stack
Video 11
-
FAM-TERRA 0 hr
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the FAM- TERRA channel at 0 hour timepoint (hr means hours).
Video 12
-
A594-RGG 0hr FRAP
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the A594- RGG channel at 0 hour timepoint (hr means hours).
Video 13
-
Cy5-dT40 0 hr
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the Cy5- dT40 channel at 0 hour timepoint (hr means hours).
Video 14
-
FAM-TERRA 8 hr
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the FAM- TERRA channel at 8 hour timepoint (hr means hours).
Video 15
-
A594-RGG 8 hr
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the A594- RGG channel at 8 hour timepoint (hr means hours).
Video 16
-
Cy5-dT40 8 hr
Description: Fluorescence recovery after photobleaching (FRAP) measurements in the Cy5- dT40 channel at 8 hour timepoint (hr means hours).
Video 17
-
VPT movie
Description: A series of images compiled as an image stack
Video 18
-
VPT movie
Description: A series of images compiled as an image stack
Video 19
-
video
Description: A series of images compiled as an image stack
Video 20
-
video.tif
Description: A series of images compiled as an image stack
Video 21
-
video.tif
Description: A series of images compiled as an image stack
>Source data for extended data (contains all the microscopy data shown in the extended data)
Dryad_source_data_for_extended_data.zip
Figure 1
- a
-
0.5mgml 0 hr.tif
-
0.5mgml 24 hr.tif
-
0.25mgml 0 hr.tif
-
0.25mgml 24 hr.tif
-
1mgml 0 hr.tif
-
1mgml 24 hr.tif
Description: Files are organized in terms of concentration expressed in terms of milligrams/milliliter (corresponds to mgml) and time point (hr means hours).
-
- b
-
0.5mg/ml 0 hr.tif
-
0.5mg/ml 24 hr.tif
-
0.25mg/ml 0 hr.tif
-
0.25mg/ml 24 hr.tif
-
1mg/ml 0 hr.tif
-
1mg/ml 24 hr.tif
Description: Files are organized in terms of concentration expressed in terms of milligrams/milliliter (corresponds to mg/ml) and time point (hr means hours).
-
Figure 2
- a
-
mut6GtoU 0 hr.tif
-
mut6GtoU 1 hr.tif
-
mut6GtoU 2 hr.tif
-
mut6GtoU 16 hr.tif
-
WT 0 hr.tif
-
WT 1 hr.tif
-
WT 2 hr.tif
-
WT 16 hr.tif
Description: Here the files are organized according to RNA variant, classified as either wild type (WT) or mutant (mut6GtoU), as well as the time point (hr means hours).
-
- b
-
0 hr.tif
-
24 hr.tif
Description: Here the files are organized in terms of time points. hr means hours.
-
Figure 3
-
A594-RGG image of FAM-TERRA and A594-RGG panel 0 hr.tif
-
A594-RGG image of FAM-TERRA and A594-RGG panel 4 hr.tif
-
A594-RGG image of FAM-TERRA and A594-RGG panel 8 hr.tif
-
Cy5-dT40 image of FAM-TERRA and Cy5-dT40 panel 0 hr.tif
-
Cy5-dT40 image of FAM-TERRA and Cy5-dT40 panel 4 hr.tif
-
Cy5-dT40 image of FAM-TERRA and Cy5-dT40 panel 8 hr.tif
-
FAM-TERRA image of FAM-TERRA and A594-RGG panel 0 hr.tif
-
FAM-TERRA image of FAM-TERRA and A594-RGG panel 4 hr.tif
-
FAM-TERRA image of FAM-TERRA and A594-RGG panel 8 hr.tif
-
FAM-TERRA image of FAM-TERRA and Cy5-dT40 panel 0 hr.tif
-
FAM-TERRA image of FAM-TERRA and Cy5-dT40 panel 4 hr.tif
-
FAM-TERRA image of FAM-TERRA and Cy5-dT40 panel 8 hr.tif
-
Merge image of FAM-TERRA and A594-RGG panel 0 hr.tif
-
Merge image of FAM-TERRA and A594-RGG panel 4 hr.tif
-
Merge image of FAM-TERRA and A594-RGG panel 8 hr.tif
-
Merge image of FAM-TERRA and Cy5-dT40 panel 0 hr.tif
-
Merge image of FAM-TERRA and Cy5-dT40 panel 4 hr.tif
-
Merge image of FAM-TERRA and Cy5-dT40 panel 8 hr.tif
Description: Here the files are organzied in terms of a fluorescence channel in a pairwise channel combination as well as the time point. For example, A594-RGG image of FAM-TERRA and A594-RGG panel 0 hr refers to A594-RGG fluorescence channel in a pairwise channel combination of FAM-TERRA and A594-RGG, at the 0 hour time point (hr means hours).
Figure 4
-
CAGx20 Cy5-dT40 0 hr.tif
-
CAGx20 Cy5-dT40 24 hr.tif
-
CAGx20 Merge 0 hr.tif
-
CAGx20 Merge 24 hr.tif
-
CAGx20 SYTO-13 0 hr.tif
-
CAGx20 SYTO-13 24 hr.tif
-
CAGx31 Cy5-dT40 0 hr.tif
-
CAGx31 Cy5-dT40 24 hr.tif
-
CAGx31 Merge 0 hr.tif
-
CAGx31 Merge 24 hr.tif
-
CAGx31 SYTO-13 0 hr.tif
-
CAGx31 SYTO-13 24 hr.tif
-
CUGx31 Cy5-dT40 0 hr.tif
-
CUGx31 Cy5-dT40 24 hr.tif
-
CUGx31 Merge 0 hr.tif
-
CUGx31 Merge 24 hr.tif
-
CUGx31 SYTO-13 0 hr.tif
-
CUGx31 SYTO-13 24 hr.tif
-
CUGx47 Cy5-dT40 0 hr.tif
-
CUGx47 Cy5-dT40 24 hr.tif
-
CUGx47 Merge 0 hr.tif
-
CUGx47 Merge 24 hr.tif
-
CUGx47 SYTO-13 0 hr.tif
-
CUGx47 SYTO-13 24 hr.tif
Description: Here the files are organized in terms of the RNA used in the sample, fluorescence channel and the time point. For example, CAGx20 Cy5-dT40 0 hr refers to a sample containing CAGx20 RNA (x20 refers to 20 RNA repeat units), Cy5-dT40 fluorescence channel and 0 hour time point (hr means hours).
Figure 5
-
A594-RGG 0 hr Bleach.tif
-
A594-RGG 0 hr Post-bleach.tif
-
A594-RGG 0 hr Pre-bleach.tif
-
A594-RGG 8 hr Bleach.tif
-
A594-RGG 8 hr Post-bleach.tif
-
A594-RGG 8 hr Pre-bleach.tif
-
Cy5-dT40 0 hr Bleach.tif
-
Cy5-dT40 0 hr Post-bleach.tif
-
Cy5-dT40 0 hr Pre-bleach.tif
-
Cy5-dT40 8 hr Bleach.tif
-
Cy5-dT40 8 hr Post-bleach.tif
-
Cy5-dT40 8 hr Pre-bleach.tif
-
FAM-TERRA 0 hr Bleach.tif
-
FAM-TERRA 0 hr Post-bleach.tif
-
FAM-TERRA 0 hr Pre-bleach.tif
-
FAM-TERRA 8 hr Bleach.tif
-
FAM-TERRA 8 hr Post-bleach.tif
-
FAM-TERRA 8 hr Pre-bleach.tif
Description: Here the files are organized in terms of fluorescence channel (A594-RGG, Cy5-dT40, or FAM-TERRA), time point (hr means hours), and assay step (Pre-bleach, Bleach, Post-bleach).
Figure 7
-
0 hr composite.tif
-
0 hr Cy5 dT40.tif
-
0 hr FAM TERRA4.tif
-
24 hr composite.tif
-
24 hr Cy5 dT40.tif
-
24 hr FAM TERRA4.tif
Description: Here the files are organized in terms of time point (0 hr or 24 hr; hr means hours) and fluorescence channel (FAM TERRA4, Cy5 dT40 or composite).
Figure 8
-
0 hr composite.tif
-
0 hr Cy5 dT40.tif
-
0 hr FAM TERRA4.tif
-
24 hr composite.tif
-
24 hr Cy5 dT40.tif
-
24 hr FAM TERRA4.tif
-
48 hr composite.tif
-
48 hr Cy5 dT40.tif
-
48 hr FAM TERRA4.tif
-
72 hr composite.tif
-
72 hr Cy5 dT40.tif
-
72 hr FAM TERRA4.tif
-
96 hr composite.tif
-
96 hr Cys dT40.tif
-
96 hr FAM TERRA4.tif
Description: Here the files are organized in terms of the time point (expressed in hours; hr means hours) and fluorescence channel (FAM TERRA4, Cy5 dT40, or composite).
Relationship between files, if important:
Additional related data collected that was not included in the current data package:
Are there multiple versions of the dataset?
If yes, name of file(s) that was updated:
Why was the file updated?
When was the file updated?
METHODOLOGICAL INFORMATION
Description of methods used for collection/generation of data: for methodological details, please see https://www.nature.com/articles/s41557-025-01847-3
Methods for processing the data: for methodological details, please see https://www.nature.com/articles/s41557-025-01847-3
Instrument- or software-specific information needed to interpret the data: Files are available in standard formats that are accessible using Fiji. For further details, please see https://www.nature.com/articles/s41557-025-01847-3
Environmental/experimental conditions: for methodological details, please see https://www.nature.com/articles/s41557-025-01847-3
People involved with sample collection, processing, analysis, and/or submission: Tharun Selvam Mahendran, Gable M Wadsworth, Anurag Singh, Ritika Gupta, and Priya R. BanerjeeThis readme file was generated on 2025-07-15 by Tharun Selvam Mahendran