Fig. 6

PKM2 nuclear translocation promotes HIF-1α-dependent glycolytic metabolism in MDSCs after C. tropicalis stimulation. A WT MDSCs were stimulated with heat-inactivated C. tropicalis (MOI = 2) for 24 h. Immunoprecipitation was carried out with antibodies against PKM2 or rabbit IgG. The immunoprecipitates and cell lysates were analyzed by immunoblotting for the indicated proteins. B, C and E WT MDSCs were stimulated with heat-inactivated C. tropicalis (MOI = 2) in the presence or absence of TEPP-46 (as indicated concentration) for 6 or 24 h. Cell lysates were analyzed by immunoblotting for the indicated proteins (B and E). Nuclear extracts were analyzed by immunoblotting for PKM2 (C). D WT MDSCs were stimulated with heat-inactivated C. tropicalis (MOI = 2) in the presence or absence of TEPP-46 (100 µM) for 6 h. Glycolytic enzymes mRNA expression were measured by q-PCR. F WT MDSCs were stimulated with C. tropicalis (MOI = 2) in the presence or absence of TEPP-46 (100 µM), then the ECAR level of these cells was measured by Agilent Seahorse XFe96 Analyzer. G and H WT MDSCs were pretransfected with PKM2 siRNA for 24 h prior to stimulation with heat-inactivated C. tropicalis (MOI = 2) for 6 or 18 h. Glycolytic enzymes mRNA expression were measured by q-PCR (G). Cell lysates were analyzed by immunoblotting for glycolytic enzymes (H). The results shown here are expressed as the mean ± SEM. Each panel is a representative experiment of at least three independent biological replicates. *p < 0.05, **p < 0.01, ***p < 0.001. The following statistical analyses were performed: unpaired Student’s t-test or one-way ANOVA where appropriate