Western Blot Analysis of Inflammasome Proteins

Protein samples extracted from renal tissues or HK-2 cells were used. SDS-PAGE and PVDF membrane transfer were performed as previously described (Wang et al., 2019 (link)). After blocking for 1 h at room temperature, the following primary antibodies were used: anti-GSDMD-N (1:1000, CST, USA), anti-cleaved caspase-1 (1:1000, CST), anti-NLRP3 (1:1000, Proteintech), anti-IL-1β (1:1000, Abcam), anti-IL-18 (1:1000, Proteintech), anti-TXNIP (1:1000, Abcam), and anti-β-actin (1:1000, Santa Cruz, USA). Secondary antibodies were used at a dilution of 1:5000. Signals were detected with a ChemiDoc XRS system (Bio-Rad, CA, USA). ImageJ software (NIH, Bethesda, MD, USA) was used to quantify the protein bands against β-actin.