Fig. 5. S1P stimulation induces MITF degradation via the ERK signaling pathway. (A) After serum starvation, Mel-Ab cells were stimulated with 10 µM of S1P at the times indicated. Cell lysates were then immunoprecipitated with antibody against ERK1/2, and the MITF level was measured by immunoblotting. Cells were stimulated with 10 µM S1P for 10 minutes (B) or for 180 minutes (C) in the absence or presence of 20 µM PD98059. Whole cell lysates were then subjected to western blot analysis with antibodies against MITF, phospho-specific ERK, tyrosinase and TRP-1. Actin antibody was used a loading control.

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