A) Phase contrast image of cultured primary wild-type RPE cells at low magnification (× 40); Bar = 20 μm. B) Protein extracts prepared from wild-type murine RPE primary cells, ashen melanocytes, melan-ink (wild type) melanocytes and HEK-293 were separated by SDS–PAGE and immunoblotted for Rab27a, Myrip, RPE65, MyoVa and calnexin (as a loading control) as described under Materials and Methods. C) Plasmids encoding V5-MyoVIIa tail, GFP-Myrip and Rab27a were co-transfected into COS-7 cells and the expressed proteins were immunoprecipitated with anti-V5 or anti-Rab27a antibody as described under Materials and Methods. The bottom panels show total lysates after transfection, indicating transfection efficiencies. Top panels show immunoprecipitated proteins revealed by the indicated antibodies.