Fig. 4

Silencing CAV1 in FLSs in vitro promotes NLRP3 inflammasome-mediated pyroptosis and is associated with synovitis. (A). qRT‒PCR of CAV1, IL-1β, IL-18, NLRP3, TNF-α, TLR4, and MMP13 in FLSs transfected with si-CAV1 for 2 days or transfected with the control (n = 6). (B). WB and quantification of CAV1, caspase-1 and NLRP3 levels after transfection with si-CAV1 for 3 days or after transfection with the control (n = 3). (C). qRT‒PCR analysis of CAV1, IL-1β, IL-18, NLRP3, TNF-α, TLR4, and MMP13 in CL-stimulated FLSs after transfection with si-CAV1 for 2 days or the control (n = 3). (D). Representative images of coimmunostaining for CAV1 and NLRP3 in FLSs with or without CL pretreatment after transfection with si-CAV1 or the control. (E). The rates of pyroptosis were analysed via flow cytometry via Annexin V-FITC/PI double staining, and the number of pyroptotic cells was determined via FACS. (F). ELISA was used to detect the concentration of IL-1β in the cell supernatants of FLSs treated with CLs and si-CAV1 or the control (n = 3). All the data are presented as the means ± SEMs. Paired t tests and one-way analysis of variance (ANOVA) were used for statistical analysis. * p < 0.05, ** p < 0.01, ***p < 0.001