Fig. 2
Glycolysis was promoted upon TNFα- and TGFβ-induced EMT. Panc-1 cells were first cultured in the presence of 40 ng/ml TNFα or 10 ng/ml TGFβ or both for 72 h to induce EMT. a Rate of glucose uptake was measured using the non-metabolisable glucose analogue 3H-2-deoxy-glucose tracer over an 8-min period. The amount of 3H radiation trapped in the cells were counted using a beta-counter. b Lactate assay was performed on cell culture media collected after 72 h from the respective groups. c Western blot detection of hexokinase II (HK2), fructose-1,6-bisphosphotase1 (FBP1), fructose-1,6-bisphosphotase 2 (FBP2), pyruvate kinase isoform 2 (PKM2) and lactate dehydrogenase-A (LDH-A). β-actin was used as a loading control. d-l qPCR measurements of SLC2A1 (encoding Glut1), SLC2A3 (encoding Glut3), lactate dehydrogenase B (LDH-B), monocarboxylate transporter 1, (MCT1), monocarboxylate transporter 4 (MCT4) and pyruvate dehydrogenase kinase 1-4 (PDK 1-4) transcripts using β-actin as a housekeeper. Results are shown as mean ± SEM with n = 5 (a) or n = 3 (b-l). *p < 0.05, **p < 0.01, ***p < 0.001