Fig. 3

Ectopic overexpression and knockdown of TKTL1 influences the Warburg effect in melanoma cells. a QRT-PCR was employed to evaluate the expression of TKTL1 in LM-MEL-59 after treatment with either TKTL1 or control siRNA for 72 h. b Western Blotting with a mouse monoclonal anti-TKTL1 antibody showed reduction in TKTL1 levels after siRNA treatment in LM-MEL-59 after 72 h. GAPDH was used a loading control. c TKTL1 expression was assessed by qRT-PCR in LM-MEL-44 cells transfected with TKTL1 pcDNA or empty control for 72 h. d Immunoblot of TKTL1 confirmed expression of TKTL1 after transfection of a TKTL1 expression vector in LM-MEL-44. Blots were probed with GAPDH as a control for loading and transfer. Glucose consumption was measured in cell free supernatant of e LM-MEL-59 cell line following treatment with TKTL1 or control siRNA for 72 h and g LM-MEL-44 cell line following over-expression of TKTL1 or empty vector control for 72 h. The production of lactate in culture supernatants was measured in f LM-MEL-59 after knockdown of TKTL1 for 72 h and h LM-MEL-44 after treatment with TKTL1 pcDNA or empty vector control for 72 h. Values are ± SD of three experiments in triplicate (**, p < 0.005, ***, p < 0.0005)