Extended Data Fig. 6: P2RX7 controls mitochondrial integrity in CD8⁺ T cells during immune responses.

a, Left, representative electron micrographs showing mitochondrial structures; right, mitochondrial area measurements of in vitro-activated wild-type and P2rx7^−/− P14 cells following culture for 72 h in IL-2 or IL-15. Representative of 3 independent experiments (n = 8–9 in total). Black bars indicate 500 nm. b, c, Wild-type and P2rx7^−/− P14 cells were activated in vitro for 72 h and assayed at that time (b) or after a further 72 h of culture in IL-2 or IL-15 (as in Fig. 3b) (c) for OCR. Graphs show values for proton leak (the difference in OCR values after oligomycin and after antimycin A/rotenone addition; Fig. 3b). Data are from three independent experiments (n = 11–18 total). d, Calculated proton leak derived from OCR measurements on in vivo activated wild-type and P2rx7^−/− P14 CD8⁺ MPECs described in Fig. 2d–f (data from three independent experiments, data pooled from five mice per experiment; n = 4–6 wells in total). e–h, Wild-type and P2rx7^−/− P14 cells were activated in vitro and polarized with either IL-2 or IL-15 (as in Fig. 3b); total cell (e–g) and mitochondrial (g, h) protein extracts were collected for protein quantification experiments. Data from three independent experiments, samples pooled from n = 6 mice total (2 mice per experiment). e, Representative blot showing Opa1 expression in polarized wild-type or P2rx7^−/− P14 cells, in comparison with β-actin (for gel source data, see Supplementary Fig. 1). f, Opa1 protein levels in polarized wild-type or P2rx7^−/− P14 cells, normalized to β-actin. g, Mitochondrial concentration (normalized by total protein concentration) in polarized wild-type or P2rx7^−/− P14 cells. h, Opa1 protein levels in polarized wild-type or P2rx7^−/− P14 cells, normalized to total mitochondrial concentration. a–d, f–h, Mean ± s.e.m.; a, b, d, two-tailed Student’s t-test; c, f–h, two-sided Mann–Whitney’s test; *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001.

Source Data