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. Author manuscript; available in PMC: 2010 Aug 1.
Published in final edited form as: Mol Cancer Ther. 2009 Aug 11;8(8):2412–2423. doi: 10.1158/1535-7163.MCT-09-0186

Figure 3.

Figure 3

Expression of shRNA specific to Mig-7 decreases S6 kinase, ERK1/2 and Akt phosphorylation in RL95 cells. (a) Normalized fluorescence intensity from analysis of phosphorylated ERK1/2, Akt and S6 kinase in RL95 shRNA 1-3 (knockdown) and 3-1 (control) stably transfected cells. Cells were plated and prepared as given in Methods. 1-3 shRNA expressing cells in which levels of Mig-7 are significantly reduced by >50% (2) showed significant phosphorylation decreases in ERK1/2 by 40% (p = 0.002), in Akt by 10% (p = 0.0449), and in S6 kinase by 30% (p = 0.0155) compared to control RL95 cells that express levels of Mig-7 similar to the RL95 parental cell line. (b) Normalized fluorescence intensities from analysis of phosphorylated PRAS40 and IGF-1R. All phosphorylation fluorescence intensities were normalized to total respective protein fluorescence intensities. These experiments were performed in triplicate.