Vacuolar transport and function of Saccharomyces cerevisiae sterol ester hydrolase Tgl1
Data files
Mar 02, 2026 version files 336.14 MB
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RawData_Figures_Revision_3.zip
336.12 MB
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README.md
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Abstract
This dataset supports the thesis titled "Vacuolar transport and function of Saccharomyces cerevisiae sterol ester hydrolase Tgl1." It includes results from microscopic analyses, thin-layer chromatography, and Western blotting of wild-type and mutant strains, as well as the corresponding statistical analyses. For fluorescence microscopy, images were acquired as single optical mid-sections at the approximate midpoint of the cells, without Z-stack collection. The vacuolar membrane was visualized using Vph1-EGFP. Vacuolar membrane microdomains were observed as discontinuous patterns of fluorescence signals on the vacuolar membrane. The localization of Tgl1 was visualized using Tgl1-mCherry. For statistical analysis, 30 cells were analyzed per field in three independent microscopic fields for each strain. Thin-layer chromatography data were also statistically analyzed based on results from three independent experiments. This dataset focus on the localization and function of Tgl1 after a diauxic shift.
Dataset DOI: 10.5061/dryad.zgmsbccqp
Description of the data and file structure
This repository contains data and analysis scripts related to the paper "Vacuolar transport and function of Saccharomyces cerevisiae sterol ester hydrolase Tgl1".
Formation of vacuolar membrane microdomains and localization of Tgl1 are observed by microscope.
Graphs represent the results from three independent experiments.
The activity of sterol ester hydrolase was detected by subjecting the reaction product of the enzyme with a radioactively labeled substrate to thin-layer chromatography.
Files and variables
File: RawData_Figures_Revision_3.zip
Description:
For microscopic observation and thin-layer chromatography, each file contains the raw data and the processed data used for the figures in this paper.
Figure 1A
Raw data from fluorescence microscopy analysis of vacuolar membrane microdomains.
The vacuolar membrane is visualized using Vph1–EGFP.
The file names include the strain name, growth phase, and cultivation time.
-1 WT log phase (8h)
-2 WT stat phase (24h)
-3 are1Δare2Δ stat phase (8h)
-4 are1Δare2Δ stat phase (24h)
Each file contains microscopy images labeled as follows.
File names consist of the date of microscopic observation, a serial number, and the imaging method.
The term “GFP” in the file name indicates that the image was acquired using GFP fluorescence channel.
The term “DIC” in the file name indicates that the image was acquired using differential interference contrast microscopy.
An exclamation mark (“!”) in the file name indicates that the image was deconvolved.
The term “crop” in the file name indicates that the image represents a cropped region of the image with the same number.
The term “300” or “300r” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The term “bar” in the file name indicates that a scale bar has been added to the corresponding image.
Figure 1B
-Figure 1B_Graphs and statistical analysis.xlsx: Contains the raw measurements from fluorescence microscopy images (Figure 1A).
For statistical analysis, 30 cells were analyzed per field in three independent microscopic fields for each strain.
Cells with vacuolar membrane microdomains were defined as those exhibiting dot-like EGFP signals and were counted as 1.
In contrast, cells without microdomains showed uniformly distributed EGFP fluorescence along the circular vacuolar membrane and were counted as 0.
Statistical analyses were performed based on these raw data.
Figure 1C
Raw data from fluorescence microscopy analysis of vacuolar membrane microdomains.
The vacuolar membrane is visualized using Vph1–EGFP.
The file names include the strain name, growth phase, and cultivation time.
-1 WT stat phase (24h)
-2 tgl1Δ stat phase (24h)
-3 yeh1Δ stat phase (24h)
-4 yeh2Δ stat phase (24h)
-5 tgl1Δyeh1Δ stat phase (24h)
-6 tgl1Δyeh2Δ stat phase (24h)
-7 yeh1Δyeh2Δ stat phase (24h)
-8 tgl1Δyeh1Δyeh2Δ stat phase (24h)
Each file contains microscopy images labeled as follows.
File names consist of the date of microscopic observation, a serial number, and the imaging method.
The term “GFP” in the file name indicates that the image was acquired using GFP fluorescence channel.
The term “DIC” in the file name indicates that the image was acquired using differential interference contrast microscopy.
The term “crop” in the file name indicates that the image represents a cropped region of the image with the same number.
The term “300” or “300r” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The term “bar” in the file name indicates that a scale bar has been added to the corresponding image.
The term “8bit” in the file name indicates that the image was converted to 8-bit from the image with the same number.
Figure 1D
-Figure 1D_Graphs and statistical analysis.xlsx: Contains the raw measurements from fluorescence microscopy images (Figure 1C).
For statistical analysis, 30 cells were analyzed per field in three independent microscopic fields for each strain.
Cells with vacuolar membrane microdomains were defined as those exhibiting dot-like EGFP signals and were counted as 1.
In contrast, cells without microdomains showed uniformly distributed EGFP fluorescence along the circular vacuolar membrane and were counted as 0.
Statistical analyses were performed based on these raw data.
Figure 2A
-Raw data and the processed data from immunoblot analysis of vacuolar protein CPY and lipid droplet-localized protein Ayr1 are included directly.
The term “crop” in the file name indicates that the image represents a cropped region of the image with the same number.
The term “300” or “300r” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The term “8bit” in the file name indicates that the image was converted to 8-bit from the image with the same number.
The term “TCL” in the file name indicates that results obtained from samples prepared from total cell lysates.
The term “Vacuole” in the file name indicates that results obtained from samples prepared from purified vacuoles by ultracentrifugation.
The term “MW marker” in the file name indicates a molecular weight marker.
Figure 2B
-Raw data and the processed data from thin-layer chromatography analysis of sterol ester are included directly.
The term “200” in the file name indicates that the images were acquired using a Typhoon™ FLA 9500 with a sensitivity setting of 200.
The term “crop” in the file name indicates that the image represents a cropped region of the image with the same number.
The term “100re” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The file named “labeled and edited images” contain images in which sample names have been added to the images within the same file.
Figure 2B_Graph and statistical analysis.xlsx contains the raw measurements from thin-layer chromatography images (Figure 2B).
This Excel file contains the raw numerical data obtained from the quantification of band intensities from multiple independent thin-layer chromatography (TLC) experiments, as well as a compiled summary table.
In the table, “Sterol ester” refers to the intensity of the upper band in the TLC image, and “FFA” refers to the intensity of the lower band.
“ProK” indicates Proteinase K treatment; “+” denotes samples treated with Proteinase K, and “−” denotes samples without Proteinase K treatment.
“Mean” represents the average band intensity within the selected region of interest (ROI), “Min” the minimum value, and “Max” the maximum value.
“IntDen” represents the integrated density (sum of pixel intensities within the ROI), whereas “RawIntDen” represents the uncorrected sum of pixel intensities.
Quantitative analyses were performed using the “IntDen” values. Ratios for each strain were calculated relative to WT, which was set to 1.**
Figure 3A
Raw data from fluorescence microscopy analysis of vacuolar membrane microdomains and localization of Tgl1-mCherry.
The vacuolar membrane is visualized using Vph1–EGFP.
The file names include the strain name and cultivation time.
-1 WT Tgl1-mCherry 24h
-2 atg1Δ Tgl1-mCherry 24h
-3 vps27Δ Tgl1-mCherry 24h
-4 vps23Δ Tgl1-mCherry 24h
-5 vps36Δ Tgl1-mCherry 24h
-6 snf7Δ Tgl1-mCherry 40h
-7 vps4Δ Tgl1-mCherry 24h
Each file contains microscopy images labeled as follows.
File names consist of the date of microscopic observation, a serial number, and the imaging method.
Files containing multiple imaging methods indicate that the images were acquired using the CellSens Process Manager.
The term “C0001” in the file name indicates that the image was acquired using differential interference contrast microscopy.
The term “C0002” in the file name indicates that the image was acquired using RFP fluorescence channel.
The term “C0003” in the file name indicates that the image was acquired using GFP fluorescence channel.
The term “crop” in the file name indicates that the image represents a cropped region of the image with the same number.
The term “300” or “300r” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The term “bar” in the file name indicates that a scale bar has been added to the corresponding image.
The term “8bit” in the file name indicates that the image was converted to 8-bit from the image with the same number.
Figure 3B
-Figure 3B_Graph and statistical analysis.xlsx contains the raw measurements from fluorescence microscopy images (Figure 3A).
For statistical analysis, 30 cells were analyzed per field in three independent microscopic fields for each strain.
Cells with vacuolar membrane microdomains were defined as those exhibiting dot-like EGFP signals and were counted as 1.
In contrast, cells without microdomains showed uniformly distributed EGFP fluorescence along the circular vacuolar membrane and were counted as 0.
Statistical analyses were performed based on these raw data.
Figure S1
-Raw data and the processed data from immunoblot analysis of lipid droplet-localized protein Erg6 and cytosol protein Actin are included directly.
This dataset serves as supplementary information confirming that the amount of lipid droplets incorporated into the vacuole is comparable between the wild-type and the tgl1Δ strain.
The vps27Δ strain was included as a negative control in this analysis.
File names include the primary antibody used for detection.
The term “300” or “300r” in the file name indicates that the image was adjusted to a resolution of ≥300 dpi from the corresponding image.
The term “8bit” in the file name indicates that the image was converted to 8-bit from the image with the same number.
The term “MW marker” in the file name indicates a molecular weight marker.
[Code/software]
Fluorescence microscopy image ware captured and processed using Olympus cellSens software.
Thin-layer chromatography results were visualized using a Typhoon™ FLA 9500 imager and processed by imageJ.
Access information
Other publicly accessible locations of the data:
Data was derived from the following sources: