Figure 2. Inhibition of BRAFV600E, expression of GLI1 and SHH-GLI pathway inhibition by NVP-LDE225 in human melanoma cells in vitro.
A) LOX IMVI and UACC 257 with PLX-4032 at the dose of 1 µM for 24 hr. subsequently protein lystaes prepared and subjected to WB analysis for the expression of GLI1 and phospho-ERK1/2. B) Effect of NVP-LDE225 on PTCH1 promoter. In total, 1 µg of PTCH1 pGL3b-hPTCH1-prom-wt or pGL3b-hPTCH1-prom-mut luciferase construct and reporter were cotransfected into LOX IMVI cells. Cells were subsequently treated with 10 µM of NVP-LDE225 or cyclopamine for 4 hours (time point selected base on kinetic experiments). Fold activation was calculated relative to cells transfected with 3 µg of pB-actin-RL. One representative experiment of 2 is shown.