Figure 4. FOXO1 and TCF1 overexpression induce chromatin remodeling at their putative DNA-binding motifs.

a-g, Bulk ATAC-sequencing analyses of day 15 tNGFR-purified CD8⁺ HA.28ζ CAR T cells overexpressing tNGFR (black), TCF1 (red), or FOXO1 (blue) (n = 3 donors). a, Principal component analysis (PCA). b, Differential open and closed peaks compared to tNGFR controls c, Chromatin accessibility tracks at IL7R, TOX, KLF3, and FASLG loci for a representative donor. d, Enrichment of an early exhausted T cell chromatin accessibility signature based on ATAC-seq data from Satpathy et al.⁹ Center line represents the mean of 3 donors. e,f, Rank ordered plot of differentially accessible transcription factor binding motifs in FOXO1_OE cells (e) and TCF1_OE cells (f) versus tNGFR controls. Transcription factor families are annotated by color. g, Heatmap and hierarchical clustering of mean differential motif accessibility. Scale shows normalized z-scores for each motif.