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. 2022 Apr 1;132(7):e155224. doi: 10.1172/JCI155224

Figure 5. SENP7-dependent reduction of PTEN controls CD8+ T cell metabolism and function.

Figure 5

(A) Immunoblot analysis of the indicated proteins in WT and KO CD8+ T cells after stimulation with anti-CD3 and anti-CD28 antibodies. (B) PI3K activity in WT and KO CD8+ T cells stimulated with anti-CD3 and anti-CD28 antibodies was analyzed using an Echelon kit (n = 3). PIP3, phosphatidylinositol (3,4,5)-trisphosphate. (C) Immunoblot analysis of the indicated proteins in WT and KO CD8+ T cells after stimulation with anti-CD3 and anti-CD28 antibodies. (D and E) Flow cytometric analysis of p-S6 (D) and Ki-67 (E) expression in WT, Senp7fl/fl Cd4-Cre (KO), Ptenfl/fl Cd4-Cre (Pten-KO), and Ptenfl/fl Senp7fl/fl Cd4-Cre (DKO) CD8+ T cells stimulated with anti-CD3 and anti-CD28 antibodies for 2 hours (D, n = 3) or 2 days (E, n = 4). (F) Flow cytometric analysis of the division of WT, KO, Pten-KO, and DKO CD8+ T cells labeled with CFSE upon stimulation with anti-CD3 and anti-CD28 antibodies for 72 hours (n = 3). (G and H) ECAR (G) and OCR (H) of WT, KO, Pten-KO, and DKO CD8+ T cells stimulated for 6 hours with anti-CD3 and anti-CD28 antibodies. (I) Tumor growth of MC38 tumor–bearing Rag1-KO mice injected with 2 × 106 WT, KO, Pten-KO, or DKO CD8+ T cells along with 2 × 106 WT CD4+ T cells on day 7 after tumor cell inoculation (n = 10). (J) Flow cytometric analysis of IFN-γ–producing CD8+ T cells in the tumors of Rag1-KO mice from I (day 12, n = 4). Data shown are representative of 3 independent experiments and are presented as the mean ± SEM. *P < 0.05 and **P < 0.01, by 1-way ANOVA with Tukey’s multiple-comparison test (D–H and J); 2-way ANOVA with Geisser-Greenhouse correction (I); and 2-tailed Student’s t test (B).