Cleaved il 1β

We detected the role of UA in cell pyrophosis. We observed that LPS stimulated the production of NLRP3, The supernatants were collected through centrifugation and subjected to protein concentration determination with bicinchoninic acid (BCA) protein assay kit (Beyotime Biotechnology, Shanghai, China). Proteins were resolved with 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), transferred onto polyvinylidene difluoride (PVDF) membranes. Membranes were then blocked with 5% bovine serum albumin (BSA) followed by incubation with primary antibodies targeting LC3-II/I (1:1000; Thermo Scientific, Waltham, MA, USA), Beclin (1:1000; Abcam, Cambridge, UK), p62 (1:1000; Abcam), gasdermin D (GSDMD, 1:1000; Abcam), GSDMD-N (1:1000; Abcam), NLR family pyrin domain containing 3 (NLRP3, 1:1000; Abcam), pro-caspase-1 (1:1000; Abcam), cleavedcaspase-1 (1:1000; Abcam), pro-IL-1β (1:1000; Abcam), cleaved IL-1β (1:1000; Abcam), activating transcription factor 6 (ATF6, 1:1000; Abcam), C/EBP-homologous protein (CHOP, 1:1000; Abcam), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH, 1:10000; Abcam). Membranes were incubated with HRP-conjugated secondary antibodies at a ratio of 1:1000 for 2 h after rinsing in tris buffered saline with tween (TBST) for 15 min. The signals were detected with enhanced chemiluminescence (ECL) detection kit.