BMP, Mitf, and RET on melanocytes and melanoma cell

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Western blotting revealed expression of the Ret protein in NCCmelb4M5 and in Mel-Ret cells, but in contrast, there was no expression of the Ret protein in NCCmelb4 or NCCmelan5 cells. Immunostaining also revealed that NCCmelb4M5 and Mel-Ret cells are positive for Ret, but NCCmelb4 and NCCmelan5 cells are negative for Ret. Thus, Ret protein is expressed in most immature melanoblasts, while melanocytes are negative for Ret. BMP-4 was added to the medium and incubated for 3 days at varying concentrations. After incubation with 10 ng/ml BMP-4 for 3 days, Ret protein expression was decreased, and disappeared completely when BMP-4 was added at 20 ng/ml. Ret protein expression in NCCmelb4M5 cells showed a dose-dependent decrease in expression elicited by BMP-4. Based on these findings, the combined effects of Ret and BMP-4 might provide important clues towards understanding the roles and working mechanisms in melanocyte development.