Fig. 4
The effect of C19 treatment on RAS activation and MAPK phosphorylation. a Representative western blot image of HCT116 and LoVo cell lines treated with vehicle and C19 (n = 3). Total protein extracts were challenged with RAF-RBD beads. The membrane was probed with antibody to pan-RAS, provided with the kit, to confirm pull down of active RAS. Total extracts were immunoblotted with the antibodies as indicated and densitometric analysis in arbitrary units. *p < 0.01 (lower panel). b, c Phosphoproteome profiling of HCT116 (b) and LoVo cells (c) treated with vehicle (upper panel) and IC50 concentration of C19 (lower panel) for 24 h. Total cell lysates from colon cancer cell lines after treatment were incubated on membranes of the phosphoproteomic human Phospho-MAPK kit as described in methods. Quantification of the pixel intensities of the signals in duplicates each from one membrane is shown. Data are depicted as percentage of control (set to 100%).