Figure 4.

SIRT6 regulates Nrf2–HK2 association. A, Western blot depicting elevated nuclear Nrf2 expression in IL-1β–treated glioma cells under glucose starvation. B, siRNA-mediated knockdown of SIRT6 prevents nuclear Nrf2 accumulation in cells treated with IL-1β under glucose starvation as demonstrated by Western blot analysis. Blots were reprobed for c23 to establish equivalent loading. C, SIRT6 positively regulates nuclear HK2–Nrf2 interaction in the presence of IL-1β in glucose-starved glioma cells. Coimmunoprecipitation shows decreased HK2–Nrf2 association in IL-1β–treated glucose-deprived cells upon siRNA-mediated knockdown of SIRT6. A confounding effect of unequal precipitation on the interaction studies was ruled out by almost equal IgG levels in each condition. Blots are representative images of three independent experiments showing similar results. NSsiRNA, nonspecific siRNA; IP, immunoprecipitation; IB, immunoblotting.