Fig. 2

Palmitoylation affects VMP1 subcellular localization. A Analysis of surface expression by biotinylation. HEK293T cells expressing WT-VMP1, MT-VMP1(C8, 192S), or (C263, 278S) treated with ethanol (NC) or 2-BP, and Sertoli cells expressing WT-VMP1 or MT-VMP1 were used for surface biotinylation assays. Total and surface proteins were analyzed by western blot using anti-flag antibody. B Sertoli cells overexpressing WT-VMP1 or MT-VMP1 were double-immunostained with anti-Flag antibody and the antibody of Na⁺/K⁺-ATPase, the marker of plasma membrane (PM). This experiment was repeated three times independently. C Statistical analysis of the colocalization rate between WT-VMP1/ MT-VMP1-flag and Na⁺/K⁺-ATPase in SCs. Values are means ± SEM from n = 3 independent experiments. *P < 0.05, the P value was determined by two-sided Student’s t-test