Western blot analysis
After treatments, the cells were lysed in RIPA buffer containing protease and phosphatase inhibitors on ice. The concentration of protein was tested using the bicinchoninic acid (BCA) protein assay. Protein samples (30 μg) were separated by 6%, 10% or 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis and were transferred to 0.22-μm PVDF membranes, followed by blocking for 2 h at room temperature with 5% dried skimmed milk in Tris-buffered saline with 0.05% Tween 20. The membranes were incubated with primary antibodies at 4 ℃ overnight, including CD36 (1:1000, proteintech, Cat#18836-1-AP, RRID:AB_10597244), TLR4 (1:1000, santa, Cat#sc-10741, RRID:AB_2240715), IκBα (1:1000, CST, Cat#4812), PCNA (1:10000, proteintech, Cat#60097-1-lg, RRID:AB_2236728), NLRP3 (1:3000, CST, Cat#15101S, RRID:AB_2722591), cleaved-IL1β (1:1000, CST, Cat#83186S, RRID:AB_2800010), cleaved-caspase-1 (1:1000,CST, Cat#4199T RRID:AB_1903916), GAPDH (1:1000, ZSGB-BIO, Cat#ta-08, RRID:AB_2747414). Subsequently, the membrane was incubated with Horseradish peroxidase (HRP)-conjugated secondary antibodies (1:8000) for 2 h at room temperature. Immunoreactivity were visualized by chemiluminescence method using ChemiDocTM MP Imaging System (Tanon, China). The protein bands were quantified using a Bio-Rad Chemi EQ densitometer and Bio-Rad Quantity One software(RRID:SCR_014280) and normalized to GAPDH or PCNA.