Fig. 2

ROS production via the FAO pathway could induce apoptosis in HCC cells treated with PA. A ROS levels in HCC cells with or without 50/100 μM PA treatment for 48 hours. N: normoxia, H: hypoxia. B ROS levels in 100 μM PA-treated or untreated HCC cells under hypoxia with or without 1 mM N-acetyl-L-cysteine (NAC) treatment for 48 hours. C Western blot analysis of cleaved caspase 3 and cleaved PARP protein expression in 200 μM PA-treated HCC cells under hypoxia with or without 5 mM NAC treatment. β-actin expression was analyzed as an internal control. D Western blot analysis of HIF-1α protein expression in HCC cells under normoxia (N) and hypoxia (H) for 12 hours. In parallel, HIF-1α expression was analyzed in HCC cells that were treated with 1 mM dimethyloxalylglycine (DMOG) under normoxia, indicated as N (DMOG+). β-actin expression was analyzed as an internal control. E Using the FAO Blue system, in vitro FAO activity was analyzed in HCC cells that were treated with PA under normoxic (DMOG-) and hypoxia mimicking (DMOG+) conditions. F Western blot analysis of CPT1A protein expression in CPT1A knockdown (KD) and scramble control (SC) HCC cells. G FAO activity in KD and SC cells with or without 100 μM PA treatment under normoxic (DMOG-) and hypoxia mimicking (DMOG+) conditions. H ROS levels in KD and SC cells under normoxia (N) and hypoxia (H) with or without PA treatment for 48 hours. I Western blot analysis of cleaved caspase 3 and cleaved PARP protein expression in KD and SC cells with or without 100 μM PA treatment under hypoxia. β-actin expression was analyzed as an internal control. Values are presented as the mean ± standard error of the mean (SEM) of three independent experiments (A, B, E, G, H). N.S.: not significant, *P < 0.05, **P < 0.01 versus control