Fig. 4
Estrogen-related receptor alpha (ERRα) interacts with HIF-1α and enhances pyroptosis resistance in an NLRP3-dependent manner. A Western blot (WB) analysis of pyroptosis-related proteins in KLE and HEC-1A cell lysates of the ovERRα group treated with CoCl2 (200 µM) for 12 h; the groups not treated with CoCl2 were used as controls. Each target protein was exposed on the same gel to ensure the same exposure conditions. B Representative SEM images of KLE and HEC-1A cells and the ovERRα group treated with CoCl2 (200 µM) for 12 h; the groups not treated with CoCl2 were used as controls. Scale: 5–30 μm. C Representative images of immunofluorescence staining of HIF-1α- and GSDMD-expressing cells per well, obtained using fluorescence microscopy after cisplatin (DDP) treatment of the ovERRα and control groups for 12 h. The groups not treated with CoCl2 were used as controls. The results represent the average of three random fields per sample. Scale: 10 µm. D-F Effect of MCC950 (10 µM) treatment on the siERRα group of KLE and HEC-1A cells measured using WB (12 h), flow-cytometry analyses (24 h), and the LDH release assay (12 h). The siERRα groups treated with DDP were included as controls. G Effect of nigericin (10 µM) treatment on the ovERRα group of KLE and HEC-1A cells measured using WB (12 h). KLE and HEC-1A cells treated with DDP were included as control groups. The dose of DDP in the above-mentioned in vitro experiments was 7 µg. The results represent the average of three experimental replicates. Data are shown as the mean ± SD. Statistical tests: Student’s t-test. *p < 0.05; **p < 0. 01; ***p < 0.001; ****p < 0.0001. Abbreviations: SEM, scanning electron microscopy