Extended Data Fig. 5: icRNA-seq analysis of RNA expression in FACS-enriched FOXP3+ cells.
From: Genome-wide CRISPR screen in human T cells reveals regulators of FOXP3
a, Schematic depicting the icRNA protocol (Methods). b, Validation of FOXP3 intracellular staining using the icRNA protocol. Data is representative of three independent trials and the antibody clone used is indicated. The schematic was adapted from ref. 51, Springer Nature. c, Gene expression levels for FOXP3 measured by icRNA-seq (n = 4 donors, mean ± s.e.m.). Statistical analysis was performed with a two-tailed unpaired Student’s t-test. d, Bioanalyzer trace depicting high-quality RNA obtained from fixed and permeabilized FACS-sorted cells. e, Scatter plot of gene expression fold changes from Perturb-seq and icRNA-seq in FOXP3+ RBPJ-knockout vs FOXP3+ NTC iTreg cells. The color is representative of the −log10 of the adjusted P-value calculated using a two-tailed Wilcoxon Rank-Sum test with Bonferroni correction for multiple testing. f, Activity scores of gene signatures associated with nTregs from the Perturb-icCITE-seq experiment. P-values by a two-tailed Wilcoxon signed-rank test.
