Extended Data Fig 2: CRISPR knockout of FOXO1 promotes an exhausted phenotype in CD4+ CD19.BBζ and in HA.28ζ CAR T cells.
a, Expression of memory- (left) and exhaustion-associated markers (right) on CD4+ CD19.BBζ CAR T cells with AAVS1 gene-editing (black) or FOXO1KO (red). Histograms show a representative donor and bar graphs show mean ± s.e.m. of 3 donors (CD8+ cells appear in Fig. 1). b, TCF1 and CD62L expression in CD4+ CD19.BBζ CAR T cells. Contour plots show a representative donor and bar graphs show mean ± s.e.m. of 3 donors (CD8+ cells appear in Fig. 1). c, Mean fluorescence intensity (MFI) of CD62L in FOXO1+ and FOXO1− gated subpopulations of CD19.BBζ CAR T cells at Day 21. d, Schematic showing CD62Llo / FOXO1KO cell negative selection strategy for RNA-sequencing experiments (Fig. 1f,g). e, Expression of memory- and exhaustion-associated markers on day 15 HA.28ζ CAR T cells f, IL-2 (left) and IFNγ (right) secretion from HA.28ζ CAR T cells in response to Nalm6 leukemia. Graphs show one representative donor (n = 2 donors). a,c, Paired two-sided student’s t-test; b, 2-way ANOVA with Bonferroni’s multiple comparisons test; f, Welch’s T-test. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.