Figure 2. Validation of MEK-regulated genes in GNAQ^Q209L/P cells.

These genes were representative of those shared between GNAQ^Q209L/P and BRAF^V600E MEK-dependent signature. A, Immunoblot analysis of cells treated with 250nM selumetinib over the time, using antibodies for the indicated proteins. Each blot is representative of at least two experiments showing same results. B, C, D, Relative mRNA levels of selected ERK output genes before and after selumetinib in three GNAQ^Q209L/P cell lines. Cells were treated with selumetinib for 8h and RT-PCR was performed using gene-specific primers for DUSP6, ETV5, SPRY2 and cyclin D1. Values were normalized with GADPH and HPRT as housekeeping genes using the ΔΔCT method. Each experiment was performed in triplicates. Bars, ± sd.