Fig. 4: DPI stimulates a sustained increase in glycolytic activity through nuclear translocation of PKM2 and transcriptional activation of c-Myc. | Nature Communications

Fig. 4: DPI stimulates a sustained increase in glycolytic activity through nuclear translocation of PKM2 and transcriptional activation of c-Myc.

From: Activation of GPR3-β-arrestin2-PKM2 pathway in Kupffer cells stimulates glycolysis and inhibits obesity and liver pathogenesis

Fig. 4: DPI stimulates a sustained increase in glycolytic activity through nuclear translocation of PKM2 and transcriptional activation of c-Myc.

a DPI-induced transcription of glycolytic genes requires PKM2. WT and Pkm2−/− BMDMs were treated with DPI (50 and 500 nM) or vehicle for 24 h. The transcript levels of Pkm2, Ldha, Hk2, and c-Myc were measured by real-time qPCR. Data were collected from two independent experiments with three biological replicates per group. Transcriptional level was normalized to β-actin first and then to DMSO control. Data are presented as the mean ± sd. b, c Induction of dimeric PKM2 by DPI. ImKCs were treated with vehicle or DPI (50 and 500 nM) for 1 h. Cell lysates were run on native PAGE gel and analyzed by Western blotting. Shown are representative data (b) and summarized data (c) quantified by ImageJ from three independent experiments. d DPI induces nuclear translocation of PKM2 by Western blotting. ImKCs were treated with vehicle or DPI (50 nM) for 6 h. Proteins from cytosolic and nuclear fractions were isolated and analyzed by anti-PKM2 Western blotting. Shown are representative data, and the numbers are average expression levels of cytosolic and nuclear PKM2 from three independent experiments. e DPI induces nuclear translocation of PKM2 by confocal microscopy. ImKCs and human primary KCs (AcceGen) were treated with vehicle or DPI (50 nM) for 24 h, stained with anti-PKM2 (green) and DAPI (red), followed by confocal imaging. Shown are representative images from two independent experiments. The boxed areas are enlarged and scale bars are indicated. f DPI stimulates transactivation of c-Myc. c-Myc luciferase reporter plasmid was transfected into WT and Pkm2−/− BMDMs. Transfected cells were treated with vehicle or DPI (50 and 500 nM) for 6 h, and luciferase activities were measured. Data are presented as the mean ± sd from two independent experiments (n = 5 biological replicates). P values were calculated by the two-sided student’s t-test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Source data are provided as a Source Data file.

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