Anti ha

The liver tissues and cell lines were lysed using radioimmunoprecipitation assay (RIPA) buffer (#P1003B, Beyotime). Approximately 30 μg of total protein was added to 8% SDS PAGE gels using Bio-Rad equipment (Shanghai, China). After separation, the proteins were transferred to PVDF membranes (#ISEQ00010, Millipore, Massachusetts, USA). The protein-loaded PVDF membranes were blocked with 5% skim milk (#A600669, Sangon), immersed in solutions of the primary antibodies at 4 °C overnight, and then incubated with Alexa Fluor 680-conjugated secondary antibodies (Thermo Fisher Scientific). Tubulin-α or GAPDH was used as an internal control. The used primary antibodies were listed (anti-NLRP3, #IMG-6668A, Novus Biologicals, Colorado, USA; anti-MARCH7, #PA5-54572, Sigma Aldrich; anti-Cleaved caspase-1, #PA5-99390, Thermo Fisher Scientific; anti-Matured IL-1β, #AF401, R&D Systems, City of Emeryville, USA; anti-GSDMD-N, #GSDMD antibody Abcam EPR 19828, Abcam, Cambridge, UK; anti-Tubulin-α, #NB100-690, Novus Biologicals; anti-GAPDH, #MA1-16757, Thermo Fisher Scientific; anti-actin-β, #NB600-501, Novus Biologicals; anti-HA, #NB600-363, Novus Biologicals; anti-Flag, #MA1-91878, Thermo Fisher Scientific; anti-Ubiquitin, #NB300-130, Novus Biologicals; anti-GST, #13-6700, Thermo Fisher Scientific; anti-His, #NBP2-61482, Novus Biologicals; anti-Myc, #2276, Cell Signaling Technology, Boston, USA).