README - Source Data for "Functional divergence of the NSs protein defines interferon antagonism and viral fitness across Bhanja virus lineages" ======================================================================================================================================================= Dataset Contents ----------------- This dataset contains the source data files for figures in the research article by Clarke et al. (2026). Files Included: 1. Clarke_et_al_2026_BHAV_Raw_Data.xlsx (2.4 MB) - Excel spreadsheet containing raw numerical data for all figures - 16 separate worksheets organised by figure panels 2. Clarke_et_al_2026_Source_Data.docx (48 KB) - Microsoft Word document providing a comprehensive index - Contains descriptions and units of measurement for each dataset - Includes methodological context and data interpretation notes 3. Clarke_et_al_2026_Blots_RAW.docx (5.3MB) - Undercropped and unadjusted blots underpinning all figures in the manuscript. Software Requirements --------------------- To access the data files, you will need: - Microsoft Excel (2010 or later) OR LibreOffice Calc OR any compatible spreadsheet application Required to open: Clarke_et_al_2026_BHAV_Raw_Data.xlsx - Microsoft Word (2010 or later) OR LibreOffice Writer OR any compatible word processor Required to open: Clarke_et_al_2026_Source_Data.docx Data Organisation and File Structure ------------------------------------ Excel File Structure (Clarke_et_al_2026_BHAV_Raw_Data.xlsx): The spreadsheet contains 16 worksheets organised using the following naming conventions: 1. MAIN FIGURES (14 worksheets): - Naming pattern: [Figure number][Panel letter] - Examples: 1A, 2B, 3A, 4A, 5A, 5C, 6A, 6B, 6D, 7A, 7C, 8B, 9, 10 - Covers data for Figures 1-10 in the main manuscript 2. SUPPLEMENTARY FIGURES (2 worksheets): - Naming pattern: S[Figure number][Panel letter] - Examples: S2A, S6 - Covers data for Supplementary Figures S2 and S6 Word Document Structure (Clarke_et_al_2026_Source_Data.docx): - Provides a comprehensive overview of data organisation - Contains descriptions of each figure panel and corresponding worksheet - Includes data descriptions and units of measurement - Includes methodological context and important experimental caveats Data Format and Content Notes ----------------------------- General Data Format: - Numerical data are presented as raw values (not formatted for display) - Worksheets are organised with samples/treatments as rows and timepoints/conditions as columns - Column headers contain experimental condition labels (e.g., "0 h", "24 h", "MOI 0.01", "A549") - Multiple biological replicates are presented in separate columns - Missing data points are represented as empty cells Common Abbreviations and Terminology: - WT: Wild-type virus (wtIG690, wtibAr2709, wtR1819, wtR1329, wtR1336) - rWT: Recombinant wild-type virus - NSsV5: NSs protein with C-terminal V5 epitope tag - MOI: Multiplicity of infection - FFU/ml: Focus-forming units per millilitre (virus titre measurement) - h p.i.: Hours post-infection - d p.i.: Days post-infection - Ct: Threshold cycle (qPCR) - IFN/IFNβ: Interferon-beta - ISRE: Interferon-stimulated response element - TBK1: TANK-binding kinase 1 - NSs: Non-structural protein - S, M, L: Small, Medium, Large viral genome segments - Rep: Replicate experiment Data Type-Specific Information: Viral Replication Data (Figs 2B, 3A, 4A, 5C): - Measurements: Viral titres (FFU/ml) - Format: Titres arranged by timepoint or cell line with multiple biological replicates - Typical range: 100-10,000,000 FFU/ml - Three biological replicates per condition Cell-Based Assays (Figs 6A, 6B, 6D): - Measurements: Viral titres or IFN activity - Format: Multiple conditions (virus strains, treatments) with biological replicates - IFN data expressed in relative units (IU/ml or fold-induction) Molecular Data (Figs 7A, 7C, 8B, 9): - Measurements: Reporter activity (luciferase), Western blot densitometry - Format: Normalised values relative to control or loading control - Data typically range 0-1000 for reporter assays, or 0-3000 for densitometry In Vivo Data (Fig 10): - Measurements: Viral RNA load (qPCR Ct values, copy numbers), body weight, clinical scores - Format: Individual mouse data by tissue type (spleen, liver, brain) - Timepoint: 7 days post-infection Special Notes: - Some worksheets contain data from multiple related panels (e.g., Fig 2B includes both A549 and A549-V+NPro cell data) - Time-course data include time points as column headers with measurements in rows below - Worksheet "1A" contains genome read coverage data (NGS read depth) rather than quantitative viral data - IG690NSsV5 expression levels are consistently lower than heterologous NSs proteins due to proteasomal degradation (see Fig S5 and text); this should be considered when interpreting Western blot band intensities Data Interpretation: - Consult the Word document (Clarke_et_al_2026_Source_Data.docx) for specific descriptions of what each dataset represents - Units of measurement are provided in the Word document for each figure panel - For questions about experimental methods or data interpretation, refer to the main manuscript Methods section - For additional context on the IG690 NSs stability findings, see Supplementary Figure S5 and the Results section Version Information: - Files were last modified: 18 May 2026 - This README corresponds to the final published version of the research article Contact Information: For questions about this dataset, please contact the corresponding author: Benjamin Brennan (ben.brennan@glasgow.ac.uk) MRC-University of Glasgow Centre for Virus Research University of Glasgow, Glasgow, UK