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

Figure 2. Effect of Sur8 on Rac and Akt activation.

Figure 2

A–C. The shCon-GFP and shSur8-GFP NIH3T3 cells were treated with EGF for 10 minutes (A), HEK293 cells were transfected with the indicated plasmids or siRNAs (siGFP or siSur8 #1 and #2) (B, C). For GTP-Rac measurement, WCLs were incubated with GST-PAK-CD and analyzed by immunoblotting with an anti-Rac1 antibody. For all other measurements, WCLs were immunoblotted against the indicated proteins. D. NIH3T3 cells were transfected with either Con-GFP or Sur8-GFP plasmids, and immunocytochemical analysis was performed using anti-p-ERK or -p-Akt antibody. Cell nuclei were counterstained with DAPI. Relative intensities of the markers stained were quantified for at least 15 different cells using NIS-Elements AR 3.1. Scale bars, 50 μm. The values are mean ± s.e.m. of three independent experiments.