Fig. 3

GSDME-mediated IEC pyroptosis increases the release of HMGB1. a IECs from Gsdme^−/− (KO) mice and wild-type (WT) littermates treated with DSS for 7 days were isolated; then, total RNA was isolated and the mRNA levels of DAMPs were determined by qPCR and normalized to GAPDH. b–g IECs from KO and WT littermates treated with DSS for 7 days were isolated, then washed with PBS containing Penicillin and Streptomycin for 3 times, and finally were plated in dishes with DMEM medium containing 10%FBS for 12 h. Cell medium was used for ELISA detection. h–j Isolated IECs from KO and WT littermates were given TNF-α (50 ng/ml) plus CHX (20 µg/ml) treatment for 12 h. h, i Culture supernatant was collected for ELISA and LDH release assays. j Representative immunoblot images of IECs. Data are shown as means ± SEM from six mice in each group. Data shown are representative of three independent experiments. NS not significant; **P < 0.01; ***P < 0.001