Fig. 3

Oxidative metabolism is with TNFα- and TGFβ-induced EMT. Panc-1 cells were cultured in the presence of 40 ng/ml TNFα or 10 ng/ml TGFβ or both for 72 h to induce EMT. a Oxygen consumption rate (OCR) was measured using the Seahorse XF24 Bioanalyzer at basal conditions. b Western blot detection of mitochondrial electron transport chain complex III, II, and I. β-actin was used as a loading control. c Glucose oxidation was measured using the U-¹⁴C-glucose tracer over a 1-h period. The amount of ¹⁴C in CO₂ originated from ¹⁴C-glucose was counted using a beta counter to give an indication of glucose oxidation rate. d Total lipid in the cells was extracted using the chloroform/methanol method following 1 h incubation in U-¹⁴C-glucose-containing media. The amount of ¹⁴C incorporated in the total lipid was quantified using a beta counter. e Western blot detection of fatty acid synthase (FAS), phospho- and total-acetyl-CoA carboxylase (P-ACC and T-ACC), and stearoyl-CoA desaturase-1 (SCD1). β-actin was used as a loading control. Results are shown as mean ± SEM with n = 5 (a) or n = 3 (b, e) or n = 4 (c, d). *p < 0.05, **p < 0.01, ***p < 0.001