Fig. 3. In vitro generated T9 CAR-T cells exhibit T_cm-like and less exhausted phenotypes.

GSEA results showing a central memory or b effector memory gene signatures of polarized Th9 or Th1 cells. c Representative flow plots and d pie charts (n = 3 donors) showing CCR7 or CD45RO expression on CAR-T cells at days 7 and 16. Cells were pre-gated for GFP⁺CD3⁺CD4⁺ or GFP⁺CD3⁺CD8⁺ T cells. e Heatmap showing the expression of exhaustion and cytolytic genes in Th9 or Th1 CAR-T cells assessed by RNASeq. f Expression levels (relative to GAPDH) of exhaustion and cytolytic genes in Th1 or Th9 (upper panels), and Tc1 or Tc9 (lower panels) CAR-T cells measured by qPCR. g Heatmap illustrating the expression of genes of transcription factors in Th9 or Th1 CAR-T cells. h Expression levels (relative to GAPDH) of transcription factor genes in Th1 or Th9 (upper panels), and Tc1 or Tc9 (lower panels) CAR-T cells measured by qPCR, f and h were performed with three biological replicates and data shown are representative of two independent experiments. Data are presented as mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001, two-sided Student’s t-test. Source data are provided as a Source Data file.