Fig. 6

a 72 h treatment of BRAF mutant cells A375 and A375VR4 with the combination TH1579 (0.2 µM) and vemurafenib (0.1 µM) leads to further loss of cell viability as measured by an MTS assay when compared with either drug alone (error bars represent mean ± SD; n = 3; *p < 0.05, Student's t test). b The loss of cell viability observed in a translates into apoptosis mediated cell death which is further enhanced by the drug combination (treatment done for 48 h) as analyzed by FACS and Annexin V staining (error bars represent mean ± SD; n = 3; *p < 0.05, Student’s t test). c Combining vemurafenib with TH1579 significantly induces ROS levels (treatment done for 3 h) when compared with either drug alone (error bars represent mean ± SD; n = 3; *p < 0.05, Student's t test). d Representative images taken by microscopy to show A375 (stably expressing dTomato) and A375VR4 (stably expressing eGFP) tumors in zebrafish disease model after 72 h drug exposure with DMSO, vemurafenib (20 µM), TH1579 (20 µM) or the combination. e Individual tumor sizes from zebrafish embryos were analyzed after lysing zebrafish and measure luminescence. Data presented as % tumor size compared with the median tumor size calculated for that group (error bars represent mean ± SD; n = 3; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, Student's t test). f, g CAV-1, AXL, and AKT are most efficiently downregulated by combination treatment following 48 h treatment. Representative western blot image (f) and densiometric quantification of the western blot (g) (data shown as mean ± SD, n = 2).