Extended Data Fig. 1: Validation of SP110 depletion.
a. Gene Set Enrichment Analysis (GSEA) from the genome-wide screen from either the genes depleted (negative normalized enrichment score) or enriched (positive normalized enrichment score) in the ppp-RNA treated condition classified according to the Molecular Signatures Database (MSigDB) hallmark gene lists with a nominative p-value < 0.05 (sample permutation test). b. Validation of SP110 specific knockdown by CRISPRi at the RNA level. Data points are technical replicates. Data is representative of n = 2 independent biological replicates. c. Validation of specific gene knockdown by CRISPRi from Fig. 1d. Data points are technical replicates. Data is representative of n = 2 independent biological replicates. d. Representative flow cytometry gating strategy. Live cells are gated on SSC-A versus FSC-A. Single cells are gated on FSC-H versus FSC-A. Fluorescent positive cells are gated on FSC-A versus fluorescent protein (mCherry, GFP, or BFP). e. Validation of SP110 knockdown at the protein level in the screening cells. NT = non-targeting guide RNA. f and g. Validation of SP110 specific knockout of selected clones in indicated cell backgrounds. c1 = clone 1. h. Competition assay in Jurkat SP110-KO cells ± IFNβ treatment and ± SP110 stable overexpression. Data points represent n = 3 individual biological replicates. i. Western blot assessing SP110 and SP100 expression in primary patient-derived T-lymphoblasts. WT = healthy donor. VODI = VODI patient #1. Bars in b, c and h denote mean ± s.d. Western blots in e, f, g and i were performed 2–3 independent times with similar results. Source numerical data and unprocessed blots are available in source data.
