Figure 8

PKR protein binding also was evoked in response to glucolipitoxicity and TNFα, which remitted the deleterious effect of TNFα on MIN6 cell proliferation. Twenty-four hours after transfection with PKR-K296R or pSG5, cells were stimulated with or without glucolipitoxicity (glucose 16.7 mmol/L and 0.05 mmol/L palmitate) or proinflammatory cytokines TNFα (80 ng/mL) for 6 h. (A) Immunocomplexes of PKR were analyzed by Western blotting with antibodies against TRAF2 and TRAF6. (B) Luciferase activity was measured and shown in fold. (C) mRNA and protein levels of c-myc were determined by qRT-PCR (upper panel) and Western blotting (down panel). After transfection, MIN6 cells were stimulated with TNFα. (D) Cell viability and (E) quantification of EdU-positive β cell of MIN6 cells were shown. β-actin was detected as an internal control. Data are means ± SEM of three separate experiments. *P < 0.05 versus control.