Partial knockdown of MPST upregulates CSE by interacting with CSE

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Partial knockdown of MPST upregulates CSE by interacting with CSE Partial knockdown of MPST upregulates CSE by interacting with CSE. (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. Partial knockdown of MPST upregulates CSE by interacting with CSE. (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 of hepatic CSE protein expression. (B) Western blot analyses of hepatic CSE expression in HFD-fed MPST+/+ and MPST+/- mice. (C) 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 CSE protein expression. (D) After transfection 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 to determine CSE protein expression. (E) Immunoblot analysis of the interaction between MPST and CSE by co-immunoprecipitation. FFA-treated L02 cells were transfected with Flag-MPST and subjected to immunoprecipitation with an anti-Flag monoclonal antibody. Co-immunoprecipitated CSE was detected with an anti-CSE antibody. The results are expressed as the mean±SD of 3–5 mice per group from two independent experiments or three independent experiments. *p<0.05, **p<0.01. CSE, cystathionine γ-lyase; 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. Meng Li et al. Gut 2018;67:2169-2180 Copyright © BMJ Publishing Group Ltd & British Society of Gastroenterology. All rights reserved.