Fig. 5

HRD1 inhibited aerobic glycolysis, growth, migration, and invasion of breast cancer cells via PFKP downregulation. MDA-MB-231 cells stably expressing HRD1 were infected with a lentivirus for PFKP for 48 h. a Western blotting was performed. b Glucose uptake was determined by measuring uptake of 2-NBDG using flow cytometry. c Glucose concentration in the medium was measured using the Amplex Red glucose/glucose oxidase assay kit. d Lactate levels in the extracellular medium and the intracellular lactate levels in the cell lysates were measured using the lactate assay kit. e ATP concentration was measured using an ATP assay kit. f Extracellular acidification rate (ECAR) was measured using Seahorse XF96 Flux Analyzer. g MDA-MB-231 cells stably expressing HRD1 were infected with a lentivirus for PFKP for 72 h, and then cell proliferation was measured using CCK-8 assays. h MDA-MB-231 cells stably expressing HRD1 were infected with a lentivirus for PFKP for 3 weeks, and then colony-formation assay were performed. i, j MDA-MB-231 cells stably expressing HRD1 were infected with a lentivirus for PFKP for 48 h, and then the migration and invasion assays were performed