Blunted CD40 response in CREBBP KAT domain point mutants. (A) Schematic of CD40 signaling in B cells. (B-C) Heat maps (B) and density plots (C) of significant changes in CREBBP loading (FDR < 0.05) after CD40L + IL-4 treatment in isogenic CREBBP WT cells, showing analogous changes in CREBBP Y1503C mutant cells. ∗∗∗Wilcoxon rank-sum P < 2.2 × 10–16. (D) Bubble plots of gene set enrichment analysis for regions with increased H3K27Ac (above) or CREBBP loading (below) after CD40L + IL-4 stimulation. (E-F) Heat maps and density plots of H3K27Ac gain (E) and EP300 loading (F) over regions with significantly increased CREBBP loading in CREBBP WT cells after CD40L + IL-4 treatment (regions from panel B). ∗∗∗Wilcoxon rank-sum P < 2.2 × 10–16. (G) RNA-seq gene set variation analysis (GSVA) scores for CD40-responsive gene sets in isogenic CREBBP WT and Y1503C RL cells with or without CD40L + IL-4 stimulation. (H) Western blot of nuclear and cytoplasmic fractions for NF-κB (p50/p105; p65) and IRF4 TFs after CD40L + IL-4 treatment. (I) Fold enrichment of TF motifs within regions of H3K27Ac gain, comparing CREBBP WT (y-axis) and Y1503C (x-axis) cells. (J) NF-κB reporter assay in CREBBP WT (green) and Y1503C (purple) cells with a dose titration of CD40L. Adjusted Student t test ∗∗P < .01; ∗∗∗P < .001.

Blunted CD40 response in CREBBP KAT domain point mutants. (A) Schematic of CD40 signaling in B cells. (B-C) Heat maps (B) and density plots (C) of significant changes in CREBBP loading (FDR < 0.05) after CD40L + IL-4 treatment in isogenic CREBBP WT cells, showing analogous changes in CREBBP Y1503C mutant cells. ∗∗∗Wilcoxon rank-sum P < 2.2 × 10–16. (D) Bubble plots of gene set enrichment analysis for regions with increased H3K27Ac (above) or CREBBP loading (below) after CD40L + IL-4 stimulation. (E-F) Heat maps and density plots of H3K27Ac gain (E) and EP300 loading (F) over regions with significantly increased CREBBP loading in CREBBP WT cells after CD40L + IL-4 treatment (regions from panel B). ∗∗∗Wilcoxon rank-sum P < 2.2 × 10–16. (G) RNA-seq gene set variation analysis (GSVA) scores for CD40-responsive gene sets in isogenic CREBBP WT and Y1503C RL cells with or without CD40L + IL-4 stimulation. (H) Western blot of nuclear and cytoplasmic fractions for NF-κB (p50/p105; p65) and IRF4 TFs after CD40L + IL-4 treatment. (I) Fold enrichment of TF motifs within regions of H3K27Ac gain, comparing CREBBP WT (y-axis) and Y1503C (x-axis) cells. (J) NF-κB reporter assay in CREBBP WT (green) and Y1503C (purple) cells with a dose titration of CD40L. Adjusted Student t test ∗∗P < .01; ∗∗∗P < .001.

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