Fig. 3: SUMO-defective HK2 increases prostate cancer cell glycolysis and decreases mitochondrial respiration and ROS.

a Glucose consumption was measured in PC3 cells with different mutant forms HK2. Endogenous HK2 was knockdown by shRNA and replaced by different mutant forms with HA tag in PC3 cells. Data are presented as mean ± SEM of three biologically independent samples. Statistical significance was determined by a two-tailed Student’s t test. b Lactate production was measured in PC3 cells with different mutant forms HK2. Data are presented as mean ± SEM of three biologically independent samples. Statistical significance was determined by a two-tailed Student’s t test. c PC3 cells as described were measured by the mitochondrial stress kit to determine extracellular acidification rate (ECAR). Data are presented as mean ± SEM of three biologically independent samples. Statistical significance was determined by a two-tailed Student’s t test. d PC3 cells as described were measured by the mitochondrial stress kit to determine oxygen consumption rate (OCR). e Intracellular and mitochondrial levels of reactive oxygen species (ROS) were measured in indicated PC3 cells by flow cytometry. Left: PC3 with the different mutant form of HK2 was loaded with ROS probe DCFH-DA, and DCF fluorescent intensity was measured by flow cytometry. Right: Cells labeled with MitoTrackerTM Red CMXRos were measured by flow cytometry to see the mitochondrial level of ROS. Experiments were performed at least twice in triplicates. Source data are provided as a Source Data file.