Figure 9.

RON function, signaling and inhibition. (A) Left, cell cycle of AGS under selected growth factor and blocking monoclonal antibody combinations. Gated events = 10,000. Right, apoptosis by annexin V (abscissa) and propidium iodide (PI, ordinate) in AGS with selected growth factor and blocking monoclonal antibody combinations. Gated cells = 20,000. (B) Top, immunoblot showing p-Tyr 4G10 and p-RON, p-MET, p-STAT3 relative to loading control with exposure to varying growth factor conditions over time. C, control. Bottom, effect on p-RON and p-AKT (odyssey blot) of pre-incubation with novel extracellular monoclonal RON blocking antibody (R Ab) at varying concentration and MSP exposure versus controls. p-AKT = red, total AKT = green. (C) Top left, viability of GEC lines with SU11274 (black), pimozide (grey) or combination (red). Bottom left, isobolograms demonstrating combination indices at ED₅₀ (red) and ED₇₅ (green). Top right, viability of GEC lines with SU11274 (black), oxaliplatin (grey) or combination (red). Bottom right, isobolograms demonstrating combination indices at ED₅₀ (red) and ED₇₅ (green). (D) Effect of SU11274 inhibition on GEC lines with growth factor combinations (1 minute exposure) on global phosphorylation (p-Tyr 4G10) and p-RON, p-MET, p-STAT3 and p-EGFR relative to loading control for MKN-45 [left, SU11274(0.2 uM)] and NCI-N87 (middle, SU11274 [2 uM]). M, MSP; H, HGF. Right, effect on global phosphorylation (p-Tyr 4G10) and p-STAT3 of treatment with MSP with or without pimozide (12 uM) over time.