Fig. 4
Inhibition of Cx32 channel function attenuated NLRP3-mediated pyroptosis in H/R injury. HK-2 cells were seeded at low density (25,000 cells/cm2, no GJ formed) and high density (125,000 cells/cm2, GJ formed) and subjected to hypoxia/reoxygenation (H/R) treatment. A Western blot analysis of NLRP3, GSDMD, GSDMD-N, IL-18, and mature IL-18 in HK-2 cells. LD: low density group; HD: high density group. *p < 0.05 vs. control group at high density; # p < 0.05 vs. H/R group at low density; & p < 0.05 vs. control group at low density. Data are means ± SEM (n = 3). B Survival rate of HK-2 cells at high density in different groups. * p < 0.05 vs. control; # p < 0.05 vs. H/R. Data are means ± SEM (n = 5). C Concentration of IL-18 and IL-1β in culture supernatant of HK-2 cells at high density in different groups. * p < 0.05 vs. control; # p < 0.05 vs. H/R. Data are means ± SEM (n = 5). D Immunofluorescence results of NLRP3 expression in HK-2 cells at high density. Scale bar = 50 μM. 2APB: cells were pretreated with 2APB (25 μM) for 1 h before the next step; siRNA: Cx32-siRNA (50 nM) transfected into cells for 48 h before the next step. Cx32-OP: the cells were transfected with plasmid-Cx32 for 48 hours to achieve overexpression of Cx32 before the next step