Figure 5

Down-regulation of cav-1 expression inhibits cytokine mixture-induced NF-κB activation. (a) INS-1 cells were transfected with CON siRNA or Cav-1 siRNA for 24 h, followed by treatment with the cytokine mixture (CM; IL-1β 20 ng/ml, TNFα 20 ng/ml). After 24 h, cell lysates were subjected to western blot analysis with specific antibodies. Actin and Lamin B1 were used as cytosolic and nuclear loading controls, respectively. (b) The densities of western blot signals were measured, and the relative expression level was normalized to that of actin, Lamin B1, and the non-phosphorylated form (IKK, IκB, and NF-κB). (c) Cells were treated as described in (a) and mRNA levels of IL-1β and iNOS was determined after 6 h incubation with the CM. Values are means ± SEM from triplicate experiments, *p < 0.05 vs. CON siRNA treated with CON, #p < 0.05 vs. CON siRNA treated with CM.