Fig. 1. PANX1 is cleaved by caspase activation in TNFα-sensitive CRC cells.

A The plasma TNFα levels in CRC patients before and after chemotherapy treatment were measured by ELISA (n = 19, p = 0.0366, paired t-test). B The response to TNFα in five CRC cell lines was evaluated by cell viability assay after 24 h of treatment (n = 3). Among the five cell lines, two (HCT15 and HCT116) were sensitive to TNFα treatment. C HCT15 and HCT116 cells were treated with TNFα (25 and 50 ng/mL) for 24 h, and cell lysates were harvested for western blotting. D HCT15 and HCT116 cells were infected with lentivirus carrying shRNA targeting TNFR1 and selected with puromycin for 3 days. The knockdown efficacy was evaluated by western blotting (n = 3). ***p < 0.001. Unpaired t-test. E HCT15^shNC and HCT15^shTNFR1 cells were treated with TNFα (50 ng/mL) for 24 h. The cleavage of caspase-8, caspase-3, and PANX1 was evaluated by immunoblotting (n = 3). Similar experiments were carried out in HCT116 cells (n = 3). ***p < 0.001. One-way ANOVA test. F HCT15 and HCT116 cells were infected with lentivirus carrying shRNA targeting TNFR2 and selected with puromycin for 3 days. The knockdown efficacy was evaluated by immunoblotting (n = 3). HCT15^shNC and HCT15^shTNFR2 cells were treated with TNFα (50 ng/mL) for 24 h. The cleavage of caspase-8, caspase-3, and PANX1 was evaluated by western blotting (n = 3). Similar experiments were carried out in HCT116 cells (n = 3). *p < 0.05 and ***p < 0.001. One-way ANOVA test. G HCT15 and HCT116 cells were treated with TNFα (50 ng/mL) and the TNFR1 inhibitor R-7050 (10 μM) for 24 h. The cleavage of caspase-8, caspase-3, and PANX1 was evaluated by western blotting.