Figure 3.

BOLD-100 rewires the signaling network of BRAFMT colorectal cancer cells. RNA-seq of BOLD-100 treated VACO432 and VT1 cells for 3 and 24 hours was performed. A, PCA plot displaying the separation between the BRAFMT VACO432 and VT1 BRAFWT clone following treatment with 24 μmol/L BOLD-100 for 3 and 24 hours. B and C, Volcano plots show the up- and downregulated genes following BOLD-100 treatment at the indicated time-points in VACO432 cells. Dashed lines on the x and y-axis indicate log₂ratio of 0.58/−0.58, and −log₁₀P value = 1.3, respectively. KEGGS pathway analysis was performed using DEG (FC: +/− 1.5 P: < 0.05) genelists for VACO432 cells treated for 3 and 24 hours with BOLD-100. D, VACO432 cells (left) were co-treated with 50 μmol/L BOLD-100 alone or combined with a panel of 61 small molecule inhibitors for 48 hours and cell viability assessed using the CTG assay. Three concentrations per drug were tested (Supplementary Table S5). Waterfall plot showing mean (n = 3) robust Z-scores (rZ) for each compound concentration used in the drug screen. Negative rZ-scores indicate agents that are sensitive to BOLD-100, and vice versa. Dashed lines on graphs indicate values of 1.0 and −1.0. rZ-scores for 2/3 concentrations for AZD6738 are highlighted in red. Positive hit from primary screen (right). BRAFMT COLO-205 and HT-29 colorectal cancer cells were treated with BOLD-100 alone or combined with AZD6738 for 72 hours and cell viability assessed using the CTG assay. Absolute cell viability is shown. Dashed line indicates 50% and 25% cell viability. Representative of three independent experiments are shown. CONT, control.