https://doi.org/10.5061/dryad.3xsj3txw1

DATASET SUMMARY

This dataset contains raw and processed experimental data supporting the manuscript
“Organ-specific redox imbalance in spinal muscular atrophy mice and partial rescue by SMN antisense oligonucleotides.”

File formats:

• TIFF files (raw Western blot images)
• XLSX spreadsheets (quantification and TBARS data)
• TXT README file

Software required:

• Any software capable of opening TIFF images
• Excel or compatible spreadsheet software for XLSX files

Reusability:

• All data needed to reproduce the figures, quantifications, and statistical analyses are included.

Overview

This dataset contains all raw and processed data supporting the findings of the manuscript.
It includes:

• Raw Western blot image files (uncropped TIFFs)
• Raw lipid peroxidation (TBARS/MDA) measurements from TECAN plate reader
• Processed TBARS values (normalized per mg protein and WT-normalized)
• Complete quantification tables for all figures and supplementary figures (ready for reproduction in GraphPad Prism)
• Full documentation of standard curve parameters, excluded points, and data-processing steps
  All files are organized into transparent, reproducible folders.

Folder Structure
of Dataset_SMA_Multiorgan_Study.zip :

A. Raw_WesternBlot_Images/

Contains the uncropped, unprocessed TIFF images for all Western blots included in the manuscript.
Contains additional Western blots used as independent experiments for statistics.
Subfolders:
Figure_2-7
Supplementary_Figures S1
Supplementary Figures S3
Supplementary Figures S5
Supplementary Figures S6
Supplementary Figures S8
Supplementary Figures S10
Supplementary Figures S11
Supplementary Figures S12
Supplementary Figures S13
Supplementary Figures S14

Each folder contains:
The raw TIFF images corresponding to each blot panel

B. LipidPeroxidation_TBARS_Raw_and_ProcessedData/

Contains the raw and processed lipid peroxidation data (MDA/TBARS assay).
Files:

B1. LipidPeroxidation_TBARS_Brain_SpinalCord_Heart_RawProcessed.xlsx

Raw TECAN absorbance values (OD 532 nm)
Standard curve wells and technical replicates
Standard curve regression parameters
Excluded concentrations with justification
Processed MDA equivalents (nmol), normalization per mg protein, final WT-normalized values
Notes describing processing steps in detail

B2. LipidPeroxidation_TBARS_Heart_ASOs_RawProcessed.xlsx

Includes all individual biological plates
Each plate contains: raw values, annotation, standard curve, processed samples
For Plate 5, both initial and corrected calculations are shown in separate sheets, together with comments explaining standard-curve linearity and replicate exclusion
A final Notes sheet documents all processing steps, formulas, and decisions

C. QuantificationData_AllFigures.xlsx

Contains the final values used to generate all graphs in the manuscript and supplementary material.
Sheets include:
Figures 2-7
Supplementary Figures S1-S14

Each sheet contains:

Final normalized values exactly as used in GraphPad Prism

Biological replicate annotations

Values provided here match exactly those used in GraphPad Prism.

Sufficient detail for full reproducibility

Processing Notes (TBARS Assays)

A complete description is also present inside each .xlsx file under the Notes tab.

3.1 Raw OD values

• Exported directly from TECAN SAFIRE II.
• Technical duplicates were measured for each well.
  3.2 Blank correction
• Blank wells (0 nmol MDA) were averaged.
• All standard and sample OD values were blank-subtracted:
• Adjusted_OD = OD_raw – OD_blank
  3.3 Standard curve
• Standard concentrations: 0, 2, 6, 10 nmol MDA.
• Some concentrations (4 and 8 nmol) were excluded when they deviated from linearity.
• Standard curve equation:
• MDA_equivalents (nmol) = slope × Adjusted_OD + intercept
• Regression parameters recorded in each plate sheet.

3.4 Sample processing
For each biological replicate:
Raw OD = average of technical replicates
Adjusted OD = Raw OD – Average blank OD
MDA equivalents (nmol) = slope × Adjusted OD + intercept
Protein correction:
protein_mg = protein_µg / 1000
MDA_normalized (nmol/mg) = MDA_equivalents / protein_mg
Dilution factor correction applied where necessary
WT normalization
Each plate's WT mean was set to 1 for inter-plate comparability:
Prism_normalized_value = MDA_normalized / WT_mean

3.5 Special note for Heart ASO Plate 5

• Both initial and corrected calculations are shown.
• The corrected version excludes one non-linear technical replicate from the standard curve.
• The exclusion was based solely on deviation from linearity and does not materially affect biological interpretation.

Western Blot Data Processing

The dataset includes only raw TIFF files, without cropping or contrast adjustments.
No filtering, resizing, or background subtraction was performed prior to saving the raw images.
All quantifications were performed in GraphPad Prism using densitometry from the main manuscript.
Western blot TIFF images are named according to the following structure:
Figure#_Panel_Target_Tissue_Genotype[_Treatment][_merged].tif
Where:
Figure#_Panel = Corresponds to the figure number and panel designation as presented in the published manuscript (e.g., Figure3, Figure2A).
Target = Protein analyzed (e.g.,GPX4, GRX2, GSTp, GSR, SMN).
Tissue = Brain, Spinal cord, Heart, Liver, or Gastrocnemius.
Genotype = WT (wild type),HET (heterozygous), SMA.
Treatment = ASO (antisense oligonucleotide treatment), included where applicable.
_merged = Composite image used for figure presentation, showing the target protein together with the corresponding loading control (e.g., vinculin or total protein stain).
All TIFF files represent uncropped, unprocessed raw Western blot images corresponding directly to the quantifications provided in the processed data files.

Reproducibility

This dataset provides everything necessary to:
Reproduce the manuscript figures
Refit TBARS standard curves
Recalculate normalized MDA values
Verify Western blot quantifications
Recreate all bar graphs and statistics

Contact

For questions regarding the dataset or analyses, please contact the corresponding author listed in the manuscript.