Figure 3: PKM2-mediated HIF1α activation is required for HMGB1 release in activated macrophages.
From: PKM2 regulates the Warburg effect and promotes HMGB1 release in sepsis
(a) RAW 264.7 cells (RAW) and BMMs were stimulated with LPS (100 ng ml−1) for 24 h. The interaction between PKM2 and HIF1α was assayed by immunoprecipitation (IP) and immune blotting (IB). (b–f) RAW 264.7 cells and BMMs were transfected with control shRNA or PKM2 shRNA for 48 h and then stimulated with LPS (100 ng ml−1) for 6–24 h. The mRNA levels of indicated genes (b–e) were assayed by real-time PCR (n=3, means±s.e.m., *P<0.05 by ANOVA LSD test). AU, arbitrary units. The protein level of HIF1α was assayed by western blotting (f). (g–j) RAW 264.7 cells were transfected with indicated shRNA for 48 h and then stimulated with LPS (100 ng ml−1) for 24 h. The mRNA levels of indicated genes were assayed by real-time PCR. In parallel, the extracellular levels of HMGB1 were assayed by enzyme-linked immunosorbant assay (n=3, means±s.e.m., *P<0.05 by analysis of variance LSD test). AU, arbitrary unit.
