FIGURE 13.

UC-773587 and UC-857993 inhibit Ras signaling and selective growth of mouse fibroblasts. UC-773587 (A) and UC-857993 (B) were tested for their ability to inhibit 3-day NIH/3T3 cell growth. Cells were grown in the presence of DMSO or indicated concentrations of UC-773587 or UC-857993 for 3 days and subsequently assayed for their growth by MTS as described under “Experimental Procedures.” C and D, NIH/3T3 cells were serum-starved for 24 h and pretreated with the indicated concentrations of UC-773587 for 16 h or UC-857993 for 2 h. Cells were stimulated with 50 ng/ml EGF for 5 min and then assayed for pan-Ras activation by GST-Raf1 effector domain pulldown experiments and activation of phospho-ERK by Western blotting. UC-773587 (E) and UC-857993 (F) were tested for their ability to inhibit 3-day wild-type or H-RasG12V-expressing MEF cell growth. Cells were grown in the presence of DMSO or indicated concentrations of UC-773587 or UC-857993 for 3 days and subsequently assayed for their growth by MTS as described under “Experimental Procedures.” Cell growth data were performed in triplicate and represent the mean ± S.E. Cell growth data are plotted as relative growth on day 3 normalized to the DMSO control. Western blot data are representative of n = 3 experiments. Statistical t test analysis was performed on data in E and F using GraphPad Prism 6 (E, p value = 0.006; F, p value = 0.037).