Fig. 2.

NVP-AEW541 treatment inhibits constitutive activation of effectors downstream of IGF1R. (A) Immunoblot assays of MCF7 cells and two fresh-frozen WT GIST biopsies with specific anti-phospho-IGF1R and anti-IGF1R antibodies. Fifty micrograms of WCE from each sample was subjected to immunoblotting. (B) MCF7 cells were serum-starved for 12 h followed by either NVP-AEW541 treatment for 12 h and/or IGF-1 stimulation for 30 min. Immunoblot assays of phospho- and total IGF1R were performed. IC₅₀ for NVP-AEW541 was quantitated based on four independent viability assays listed. (C) Immunoblot assays of phospho- and total IGF1R. GIST 882 cells were serum-starved for 12 h followed by either NVP-AEW541 treatment for 12 h and/or IGF-1 stimulation for 30 min. (D) GIST-T1 cells and GIST 882 cells were treated with NVP-AEW541 for 6 h at the indicated concentrations. Equal amounts (40 μg) of WCE from each sample were subjected to immunoblotting with specific antibodies as indicated. In all cases, actin served as a loading control. IC₅₀ for NVP-AEW541 was quantitated based on four independent viability assays for each cell line.