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. 2015 Sep 5;6(32):33091–33105. doi: 10.18632/oncotarget.5173

Figure 3. Effects of Ras-ERK or PI3K pathway inhibition on Sur8-mediated Rac activation and cell migration.

Figure 3

A, B. WCLs of HEK293 cells transfected with the indicated plasmids and treated with either U0126 or LY294002 for 1 hour (A) or 15 hours (B) were incubated with GST-PAK-CD. GTP-Rac was measured as described in Figure 2. WCL aliquots were immunoblotted using the indicated antibodies. Cells in (A) were treated with EGF for 10 minutes. C. NIH3T3 cells were transfected with Sur8-GFP and treated with U0126 or LY294002. Real-time imaging was performed to monitor cell migration and the migratory paths were recorded and quantified from three independent experiments using NIS-Elements AR 3.1, values are mean ± s.e.m. Scale bars, 50 μm. D. NIH3T3 cells co-expressing Sur8-GFP and Con-RFP; or Sur8-GFP and DN-Rac-RFP were imaged to monitor cell migration and the migratory paths were recorded and quantified from three independent experiments using NIS-Elements AR 3.1, values are mean ± s.e.m. Scale bars, 50 μm.