CCG-203971 regulates cellular localization and blocks target gene expression of MRTF. A, SK-Mel-147 cells were cotransfected with RhoC and the SRE.L reporter.

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CCG-203971 regulates cellular localization and blocks target gene expression of MRTF. A, SK-Mel-147 cells were cotransfected with RhoC and the SRE.L reporter plasmid as described in Materials and Methods. CCG-203971 regulates cellular localization and blocks target gene expression of MRTF. A, SK-Mel-147 cells were cotransfected with RhoC and the SRE.L reporter plasmid as described in Materials and Methods. After transfection, cells were treated with the indicated concentrations of CCG-203971 overnight before lysis and reading luminescence in the plate reader. Data are graphed as a percentage of the DMSO control. B, Immunocytochemistry was performed on SK-Mel-147 cells which under starved conditions (0.5% FBS, 24 hours) maintained nuclear localization of MRTF-A. Individual cells were assessed for predominately nuclear (N), cytosolic (C), or an even distribution (N/C) of MRTF-A. Data are the averages of three independent experiments with at least 100 cells counted for each condition. Scale bar shown represents 200 μm. C, CCG-203971 effect on gene expression of CTGF, MYL9, and CYR61. SK-Mel-147 cells were starved in 0.5% FBS containing media for 24 hours in the presence of the indicated concentration of CCG-203971 or DMSO before isolation of RNA for qPCR analysis. Results are expressed as the mean (±SEM) of triplicate experiments (*, P < 0.05, **, P < 0.01 vs. DMSO control). Andrew J. Haak et al. Mol Cancer Ther 2017;16:193-204 ©2017 by American Association for Cancer Research