Fig. 3

NUPR1 inhibits ROS generation and oxidative stress via CYPs in HCC cells. a KEGG enrichment analysis of differentially expressed genes between NUPR1-overexpressing and control MHCC-97H cells showed that the metabolic pathway mediated by cytochrome P450 was downregulated in LV-NUPR1 cells. b The mRNA levels of genes included in CYP superfamily in MHCC-97H/MHCC-97L cells transfected with LV-NUPR1 or LV-NC were analyzed by qRT-PCR. Individual RNA values were normalized to β-actin values. c Western blot analysis was used to detect the protein expression of CYP1A1, CYP1B1, and CYP3A4 in NUPR1 overexpressing or knockdown cell lines treated with or without IR (8 Gy). d, e ROS levels and NADPH/NADP⁺ ratio in stable NUPR1 overexpressing or knockdown cell lines were measured after exposure to 8 Gy of IR. f Bar graphs show the relative levels of apoptotic cells after IR treatment (8 Gy) by staining with annexin V and DAPI in indicated cells. g Western blot was used to detect the protein levels of total/cleaved caspase-3 and total/cleaved PARP in different NUPR1 expressing cell lines treated with or without IR (8 Gy). Data are the mean of biological triplicates and are shown as the mean ± SD. P values: *P < 0.05; **P < 0.01; ***P < 0.001 and ns, not significant by Student’s t-test