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. 2022 Jul 4;18(11):4341–4356. doi: 10.7150/ijbs.71134

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Activation of VIPR1 by VIP decreases CAD phosphorylation by regulating ASS1 expression in an mTOR/p70S6K pathway dependent manner. (A) HCC cell lines HepG2 and Huh7 were transfected with VIPR1 and treated with VIP in vitro, cellular extracts were subjected to Western blotting analyses. Independent experiments were conducted for three times. (B) VIP treatment alone (200nM for 48hrs; shown as VIP) or VIP/VIPR1 activation (200nM VIP treated VIPR1 plasmid transinfected cell for 48hrs; shown as VIP/VIPR1) in both negative control siRNA (NC siRNA) and siASS1 transinfected Huh-7 cells, then Western Blot was performed to measure protein level change of phosphorylated CAD (p-CAD) as compared to the control Huh-7 (Ctrl). (C) p-CAD/CAD ratio and p-p70S6K/ p70S6K ratio changes in NC siRNA and siASS1 transfected HepG2 and Huh-7 cells after rapamycin treatment in vitro (200nM; 2 hours). Independent experiments were conducted for three times. Unpaired Student's t test was performed in panels A and C. Values represent means±SEM. *P< 0.05, **P< 0.01.