Fig. 1.

Sarcolemmal NHE1 trafficking is DNM2 activity dependent in ARVMs. (A) The transfection efficiency and overexpression of DNM2^WT or DNM2^K44A in ARVMs were detected by fluorescent microscopy and western blotting. Left: microscopic imaging of adenovirus-mediated GFP expression. Right: western bloting analysis of DNM2^WT and DNM2^K44A overexpression. GAPDH was used as a loading control. (B) Overexpression of DNM2^WT or DNM2^K44A induced a decrease or an increase in NHE1 protein expression in the cell surface respectively. A typical example of a western blot (upper panel) and the pooled data (lower panel) are shown. (C) A representative example of co-immunoprecipitation experiments for DNM2 and NHE1 proteins from freshly isolated ARVMs. Cell lysates were incubated with anti-NHE1, -DNM2 or -IgG (as a control) antibodies. The precipitates were then immunoblotted with the antibodies of DNM2 or NHE1. Data are expressed as means ± SEM of five independent experiments. *P < 0.05 compared with mock; #P < 0.05 compared with groups other than mock. S-NHE1: surface NHE1; T-NHE1: total NHE1; IB: immunoblot; IP: immunoprecipitation.