Glycolysis Evaluation by Glucose and Lactate

Glycolysis levels were evaluated by measuring glucose consumption and lactate production. The procedures were included as following. PGK1-overexpressing or PGK1-knockdown 786-O cells were seeded into 6-well plates at a same density of 1×10⁶ cells in 2.5 mL complete culture medium per well to culture for 24 h, then cell medium was collected to measure glucose consumption and lactate production level. At the same time, cells were collected to count to normalize the cell numbers between the experimental 786-O^-PGK1 (786-O^-shPGK1) and the control 786-O^-NC (786-O^-shNC) cells, which aimed to exclude the disturbance caused by cell growth difference on the relative level of glucose consumption and lactate production.
The concentrations of glucose consumption and lactate production were measured using a glucose assay kit (E1010, Applygen) and a lactate detection kit (A019, Nanjing Jiancheng Biotech) respectively, which were normalized by the cell numbers as described previously [26 (link), 27 (link)].