Figure 4
From: Combination of pan-RAF and MEK inhibitors in NRAS mutant melanoma
Negative growth in resistant cell lines upon knockdown of cyclin D3 and blockade of the MAPK pathway. A) Bar graphs of cyclin D1 and cyclin D3 RNA expression in M207 and M296 cell lines determined by transcription microarray analysis as it was described before. Different colors on the bars are the representation of signals from different probes for the same gene. B) Western blot analysis of cyclin D1 and D3 expressions in three resistant (M202, M207, M311) and three sensitive (M243, M296, MSKMEL173) cell lines treated for 24 hours with 500 nM of PRi, 25 nM of MEKi or their combination for 24 hours. C) Western blot analysis indicating successful blockade of cyclin D3 expression 72 hours after the transfection of the cells with a cyclin D3 specific siRNA pool. D) Effect of cyclin D3 knockdown on growth of resistant and sensitive cell line. In this experiment, kinetics of growth was monitored and recorded by an InCucyteZOOM device during a period of 110 hours. In the resistant cell lines (upper panel), negative growth was achieved by the combination of cyclin D3 knockdown + PRi + MEKi. All the conditions were in triplicates and the assay was repeated twice.