Figure 5

Computationally derived EDNRB network components impact EDNRB signaling through CREB phosphorylation. (a) Novel EDNRB network components impact both tyrosinase and MITF expression. MNT-1 melanoma cells are transfected with pooled siRNAs and relative MITF (black bar) and Tyrosinase (gray bar) expression is measured by quantitative RT-PCR 96 hours after transfection. *, p< 0.05. **, p< 0.01. (b) Novel components of the EDNRB network impact CREB phosphorylation. Lysates from MNT-1 cells treated with the indicated pooled siRNAs are subjected to immunoblotting with the indicated antibodies. (c) PLCD1 impacts MITF protein levels in melanocytes. Lysates from PLCD1 siRNA transfected melanocytes are subjected to immunoblotting with the indicated antibodies. (d) PLCD1 impacts CREB phophorylation in normal human melanocytes. Lysates from melanocytes transfected with the indicated siRNAs followed by EDNRB stimulation (treatment with 10 mM Endothelin-1 for 5 minutes) are subjected to immunoblotting with the indicated antibodies.