SREBP-1 pathway is involved in MPST-regulated fat accumulation in hepatocytes. SREBP-1 pathway is involved in MPST-regulated fat accumulation in hepatocytes.

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SREBP-1 pathway is involved in MPST-regulated fat accumulation in hepatocytes. SREBP-1 pathway is involved in MPST-regulated fat accumulation in hepatocytes. (A) HFD-fed C57/BL6 mice were given 1×109 plaque-forming units of AD-GFP or AD-shMPST via the tail vein every 2 weeks from the beginning of experiment and were sacrificed after 8 weeks of HFD feeding. Liver tissues were subjected to western blot analyses. (B) After transfection with scrambled siRNA (negative control) and MPST siRNA for 24 hours, L02 cells were treated with FFAs (1 mM) for 24 hours. Western blot analyses were performed to determine SREBP-1, FAS and ACC protein expressions. (C) After transfecting with plasmid containing GFP with or without full-length MPST DNA for 24 hours, L02 cells were treated with FFAs (1 mM) for 24 hours. Western blot analyses were performed. (D) After the transfection with NC and MPST siRNA for 24 hours, L02 cells were transfected with or without CSE siRNA for 24 hours. Then the cells were treated with FFAs (1 mM) for another 24 hours and subjected to western blot analyses. (E) Immunoblot analysis of the interaction between MPST and SREBP-1 by co-immunoprecipitation. FFA-treated L02 cells were transfected with Flag-MPST and subjected to immunoprecipitation with an anti-Flag monoclonal antibody. Co-immunoprecipitated SREBP-1 was detected with an anti-SREBP-1 antibody. The results are expressed as the mean±SD of five mice per group or three independent experiments. *p<0.05, **p<0.01, ***p<0.001. ACC, acetyl-CoA carboxylase; CSE, cystathionine γ-lyase; FAS, fatty acid synthase; FFA, free fatty acid; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; GFP, green fluorescent protein; HFD, high fat diet; IB, immunoblot; IP, immunoprecipitation; MPST, 3-mercaptopyruvate sulfurtransferase; siRNA, small interfering RNA; SREBP-1, sterol regulatory element-binding protein 1c. Meng Li et al. Gut 2018;67:2169-2180 Copyright © BMJ Publishing Group Ltd & British Society of Gastroenterology. All rights reserved.