Fig 1: VAMP2 is involved in EA-regulated FPN translocation.(A) mRNA expression levels of Vamp (Vamp2, Vamp8 and Vamp8), SNAP23 and syntaxin 4 in primary HSCs from C57BL/6 mice and LX-2 cells. n = 4. (B) Protein expression of VAMP2, VAMP3, VAMP8, syntaxin 4, and SNAP23 in primary HSCs and LX-2 cells treated with TGF-ß1 and/or EA. n = 5. (C) VAMP2 expression in primary HSCs and LX2 cells pretreated with MG-132 (10 µM) or CQ (1 µM) for 30 min followed by TGF-ß1 and/or EA treatment. n = 5. (D) Following TGF-ß1 treatment, VAMP2 expression was analyzed in primary HSCs and LX2 cells pretreated with CHX (100 µg/mL) for the indicated durations. n = 5. (E) Immunoblotting analysis of syntaxin 4 and SNAP23 in VAMP2 immunoprecipitates from primary HSCs and LX2 cells treated with TGF-ß1 and/or EA. n = 4. (F) Primary HSCs and LX-2 cells were infected with different concentrations of AdshVAMP2, AdVAMP2 or control adenovirus (AdLacz) for 24 h. VAMP2 protein expression was examined. n = 5. (G) After HSCs were infected with AdshVAMP2 or AdVAMP2 followed by TGF-ß1 treatment, the cells were incubated in EA for 24 h, and then the distribution of FPN and (H) FTH1 expression in those cells was examined by western blotting. n = 5. Data were normalized to GAPDH and are expressed as the mean ± SEM of three independent experiments. *P < 0.05, **P < 0.01 between the indicated groups.