Figure 6

TRAF2 plays a critical role in PKR protein-binding-induced NF-κB activation and proliferation in MIN6 cells. MIN6 cells were transfected with si-NC, si-TRAF2 or si-TRAF6 before GyrB-PKR-K296H plasmids. After 48 h of gene silencing, cells were treated with coumermycin as indicated. (A) Luciferase activity was measured and shown in fold. mRNA and protein levels of c-myc were determined by (B) qRT-PCR and (C) Western blotting. Antibodies against TRAF2 and TRAF6 were used to examine the protein levels after gene silencing. (D) Quantification of EdU-positive β cell was shown. Flow cytometric assays were carried out, with percentages of MIN6 cells at the (E) G1 phase and (F) S phase displayed. β-actin was detected as an internal control. Data are means ± SEM of three separate experiments. *P < 0.05 versus control.