Fig. 6. Effect of overexpression of N/S and A₂G₂ on myoblast fusion and colocalization of MARCKS with filamentous actin. Myoblasts cultured for 24 hours were transfected with the pcDNAs for Myc-tagged, wild-type MARCKS (A-C), N/S (D-F) and A₂G₂ (G-I). These cells were further cultured for the next 48 hours, followed by immunostaining with anti-Myc antibody and then with FITC-conjugated anti-IgG antibody, as described in Materials and Methods. They were also subjected to rhodamine-phalloidin staining (C,F,I). The middle panels (B,E,H) show the overlapped views of the left and right panels for colocalization of MARCKS and F-actin. The extents of membrane fusion were 82%, 93% and 27% for the cells transfected with the pcDNAs for Myc-tagged wild-type MARCKS, N/S and A₂G₂, respectively. Magnification, 400x (A-F); 500x (G-I).