Fig. 4

HK2 knockdown in Huh7 cells reduces glycolysis flux to lactate. a Glucose and glutamine uptake rates and lactate secretion rate in Huh7 cells expressing HK2-targeting shRNA (HK2), or non-targeting shRNA (Nt), under induced conditions (+Dox) or non-induced conditions (−Dox) (n = 8, biological replicates). b Uptake and secretion rates of pyruvate and amino acids (n = 8, biological replicates). c Mass isotopomer analysis of extracellular lactate in cultures with [1,2-¹³C]glucose and unlabeled glutamine (n = 2, biological replicates). d Mass isotopomer analysis of citrate in cells cultured with [U-¹³C]glutamine and unlabeled glucose (n = 2, biological replicates). e Mass isotopomer analysis of malate in cells cultured with [U-¹³C]glutamine and unlabeled glucose (n = 2, biological replicates). f Mass isotopomer analysis of extracellular lactate in cultures with [U-¹³C]glutamine and unlabeled glucose (n = 2, biological replicates). g Predominant metabolic pathways of glucose and glutamine metabolism in mammalian cells. h Metabolic flux distributions in Huh7 cells (HK2 - Dox) and Huh7 cells with HK2 knockdown (HK2 + Dox). Fluxes were determined by integrating uptake and secretion rates and isotopic labeling data from [1,2-¹³C]glucose and [U-¹³C]glutamine tracer experiments by metabolic flux analysis. Arrow widths indicate absolute magnitudes of net fluxes (nmol/10⁶ cells/h). Complete mass isotopomer distributions are shown in the Supplementary Data. All data represent the mean ± SD, **p < 0.001 by Welch’s unequal variances t-test