Fig. 4
From: Celastrol mitigates inflammation in sepsis by inhibiting the PKM2-dependent Warburg effect
Celastrol (Cel) directly targets PKM2 and HMGB1. a Western blotting of pull-downs to verify Cel binding to PKM2, HMGB1 and LDHA in situ. b Immunofluorescence staining of PKM2 and HMGB1 (green) and Cel-P click-conjugated to a TAMRA dye (red). Scale bar = 10 μm or 12 μm. c Surface plasmon resonance studies of the binding affinity of Cel with recombinant human PKM2, HMGB1 and LDHA. d Cellular thermal shift assays to verify Cel binding to PKM2. e Cellular thermal shift assays to verify Cel binding to HMGB1 (n = 3). *P < 0.05, **P < 0.01 vs. 37 °C. Cel-P celastrol-probe, PKM2 pyruvate kinase M2, HMGB1 high mobility group box 1, LDHA lactate dehydrogenase A