Figure 2: Upregulated PKM2 promotes aerobic glycolysis and HMGB1 release in activated macrophages.

(a–d) RAW 264.7 cells were transfected with or without indicated shRNA for 48 h and then stimulated with LPS (100 ng ml⁻¹) for 6–24 h. The level of indicated protein (a), mRNA (b), ECAR (c) and HMGB1 release (d) were assayed as described in the Methods (n=3, means±s.e.m., *P<0.05 by analysis of variance (ANOVA) LSD test). AU, arbitrary units. (e) RAW 264.7 cells were pretreated with shikonin (5 μM) for 3 h and then stimulated with LPS (100 ng ml⁻¹), TNF-α (5 ng ml⁻¹) and IFN-γ (10 ng ml⁻¹) for 24 h. HMGB1 release was assayed by enzyme-linked immunosorbant assay (n=3, means±s.e.m., *P<0.05 by ANOVA LSD test). (f,g) BMMs were transfected with or without indicated shRNA for 48 h and then stimulated with LPS (100 ng ml⁻¹), TNF-α (5 ng ml⁻¹) and IFN-γ (10 ng ml⁻¹) for 24 h. The mRNA levels of PKM2 (f) and HMGB1 release (g) were assayed (n=3, means±s.e.m., *P<0.05 by ANOVA LSD test). AU, arbitrary units.