Extended Data Fig. 6: PFKL-mediated PLIN2 S159 phosphorylation promotes the survival and proliferation of HCC cells and tumor growth.
a, Huh7 cells (2 × 104 cells) with depleted endogenous PLIN2 or PFKL and reconstituted expression of the indicated Flag-rPLIN2 (left) or Flag-rPFKL (right) were treated with or without 2-DG (25 mM) or Etomoxir (Eto, 40 μM) for 48 h. The cells were counted. Data are presented as means ± SD of three biologically independent replicates, analyzed by one-way ANOVA; n = 5. b-e, Huh7 cells with depleted endogenous PLIN2 or PFKL and reconstituted expression of the indicated Flag-rPLIN2 or Flag-rPFKL were subcutaneously injected into six-week-old male athymic BALB/c nude mice (n = 12 per group). Ten days after tumor cell injection, 0.2 mL of 2-DG (500 mg/kg) or PBS control was intraperitoneally injected daily for 18 days (n = 6). The tumor volumes (b) and weights (c) were determined. Data are presented as means ± SD of 6 biologically independent replicates, analyzed by one-way ANOVA. The indicated tumor tissues were analyzed by TUNEL assay. Representative images from three independent experiments are shown. Apoptotic cells in 10 microscope fields were quantified. Data were analyzed by two-sided unpaired Student’s t-test; n = 10 (d, e).
