Fig. 6: ME2fl assembles a glycolytic complex in the cytosol.
From: AKT1 phosphorylation of cytoplasmic ME2 induces a metabolic switch to glycolysis for tumorigenesis
a–c Cytoplasmic fractions of H1299 cells expressing vector control shRNA (shCtrl) or shRNA targeting ME2 (shME2) were immunoprecipitated with anti-LDHA (a), anti-PKM2 (b) and anti-GAPDH (c) antibodies. The immunoprecipitants were analyzed by western blotting using the antibodies against ME2fl, PFKL, PKM2, GAPDH, or LDHA as indicated. d Confocal imaging of shCtrl or shME2 H1299 cells comparing staining between GAPDH and PFKL, PKM2, LDHA or ME2. Scale bars, 10 μm. e Protein complexes from shCtrl or shME2 H1299 cells were separated by sucrose density gradient centrifugation and analyzed by western blotting with the indicated antibodies (left panel). The data were quantified using Image J v.1.46 (right panel). All data are representative of three independent experiments. f Lysis from prostate tissues of the wild type (Pb-Cre-PtenL/L) and Pten-condition knockout (Pb-Cre+PtenL/L) mice were immunoprecipitated using an anti-ME2 antibody. Whole-cell lysis (input) and the immunoprecipitations were analyzed by western blot with the indicated antibodies. All data are representative of three independent experiments.
