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. 2023 Jan 23;220(4):e20220686. doi: 10.1084/jem.20220686

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© 2023 Sun et al.

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Figure S2. — STAT3 promotes the development of terminally differentiated CD8⁺ T cells. (A) Numbers of T_ex^prog (upper panel) and T_ex^term (bottom panel) OT-I cells from B16-OVA tumors on day 14 and day 20 after tumor inoculation. (B) Representative plots (upper panel) and quantifications (bottom panel) of Ly108 and TIM-3 expression on OT-I cells from TDLNs in B16-OVA tumor model on day 20 after tumor inoculation. (C) Representative plots (left panel) and quantifications (right panel) of Bcl6 and Blimp1 expression in OT-I cells from B16-OVA tumors on day 14 and day 20 after tumor inoculation. (D) Representative plots (left panel) and quantifications (right panel) of TCF1 and CX3CR1 expression in/on OT-I cells from B16-OVA tumors on day 20 after tumor inoculation. (E) Representative plots (upper panel) and quantifications (bottom panel) of TOX and T-bet expression in OT-I cells from B16-OVA tumors on day 20 after tumor inoculation. (F) GSEA results for identifying gene signatures of WT and Stat3^−/− OT-I TILs compared with other T cell subsets (GEO accession nos. GSE65660 and GSE8678). NES, normalized enrichment score; FDR, false discovery rate q-value. (G) Volcanic plot showing DEGs in WT and Stat3^−/− OT-I TILs. (H) Heatmap illustrating the relative expression of signature genes related to cell cycle pathway in WT and Stat3^−/− OT-I TILs. (I) GSEA result for identifying gene signatures of WT and Stat3^−/− OT-I TILs compared to OXPHOS feature genes (GSEA: MM3893). NES, normalized enrichment score; FDR, false discovery rate q-value. (J) Heatmap illustrating the relative expression of signature genes related to OXPHOS pathway in WT and Stat3^−/− OT-I TILs. (K) Representative plots (left panel) and quantification (right panel) of T-bet expression in splenic OT-I cells on day 8 after LM-OVA infection. (L) Representative plots (left panel) and quantifications (right panel) of CD127 and KLRG1 expression on splenic OT-I cells on day 8 after LM-OVA infection. (M) Representative plots (left panel) and quantification (right panel) of Granzyme B expression in TCF1⁻Granzyme B⁺ splenic OT-I cells on day 8 after LM-OVA infection. (N) OT-I cell frequencies among total splenic CD8+T cells on day 8 after LM-OVA infection. (O) Representative plots (left panel) and quantification (right panel) of Ki-67 expression in splenic OT-I cells on day 8 after LM-OVA infection. Data are representative of two (K–O) or three (A–E) independent experiments. Data are shown as mean ± SEM. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 by paired two-tailed Student’s t test (A–E and K–O).