Fig. 1: A20 is linked to glucose metabolism in HCC cells.

a A20 suppresses proliferation in Huh7 and LM3 cells. Cell proliferation was determined by cell counting. The two-tailed Student’s t-test was used. b A20 decreases HCC tumor growth in vivo. Huh7 cells stably expressing A20 and empty vector counterpart cells were subcutaneously injected into the bilateral flanks of nude mice. At 4 weeks after injection, tumors from 10 mice were extracted and photographed (left panel). Tumor diameters were measured at the indicated time points, and tumor volumes were calculated (right panel). Significant differences were evaluated by t-test. c A20 suppresses the migration of Huh7 and LM3 cells. Huh7 and LM3 cells were transfected with pcDNA3-A20 or pMKO-shA20 plasmids, and cell migration was analyzed by Transwell experiments. d Quantitative analysis of cell migration was performed by ImageJ. The numbers of migrated cells (mean ± S.D.) from three independent experiments. e, f A20 suppresses cell glucose uptake and lactate production. pcDNA3-A20 or pMKO-shA20 plasmids was transfected in Huh7 and LM3 cells, respectively, and cellular glucose uptake and lactate production were detected via glucose uptake assay and lactate colorimetric assay, respectively. Error bars represent ± S.D. for triplicate experiments. g A20 inhibits cell glycolysis. The real-time assessment of the extracellular acidification rate (ECAR) in cultured cells was examined by Seahorse XFe96 analyzer. h Relative glycolytic capacity was normalized to the cell number (means ± S.D., n = 3). (The two-tailed Student’s t-test was used. The symbol * shows statistically significant differences with *p < 0.05, **p < 0.01, and ***p < 0.001).