Increase of IRF-1 and IL-15 expression in Piezo2 KO tumor cells upon radiation depended on JAK2/STAT1 pathway. (A) The expression of p-JAK2 and p-STAT1 shown by western blot (left) and quantification (right) in WT and Piezo2^−/− MC38 with or without IR treatment for 15 min after IR were represented from three independent experiments. (B) C57BL/6 were transplanted subcutaneously with 2 × 10⁶ WT and Piezo2^−/− MC38-tdTomato⁺ cells. Tumors were treated locally with one fraction of 18-Gy IR. On day 5 after radiation, p-STAT1 expression in MC38-tdTomato⁺ cells was analyzed by flow cytometry. Representative data and quantification of p-STAT1 expression in MC38-tdTomato⁺ cells from irradiated tumors were shown from two independent experiments (n = 5–6 mice per group). (C and D) Representative data and quantification of IL-15 expression in WT and Piezo2^−/− MC38 (C) and B16F1 (D) cells at 15 min after radiation with or without JAK inhibitor (1 µM) 1 h ahead of IR were shown from three independent experiments. (E and F) Representative data and quantification of IRF-1 expression in WT and Piezo2^−/− MC38 (E) and B16F1 (F) cells at 15 min after radiation with or without JAK inhibitor (1 µM) 1 h ahead of IR were shown from three independent experiments. (G) Representative data and quantification of IFN-γ expression in WT and Piezo2^−/− MC38-tdTomato⁺ cells from tumors on day 5 upon radiation were shown from two independent experiments (n = 4–6 mice per groups). (H) Representative data and quantification of IL-15 expression in irradiated MC38 cells with or without anti-IFN-γ treatment at 100 µg/ml for 60 h are shown from two independent experiments. Data were represented as means ± SEM. Statistical analysis was performed by one-way ANOVA with multiple comparison tests (A–H). *P < 0.05; **P < 0.01; ***P < 0.001; ****P <0.0001; ns, no significant difference. Source data are available for this figure: SourceData F5.