LyGDI influences Rac membrane localizaation.
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(A) 1×107 PBM were transfected with either scrambled or LyGDI siRNA. Western blotting was done to measure LyGDI after 48 hours (upper panel). The membrane was reprobed with anti-actin to ensure equivalent loading (lower panel). Numbers in the upper panel are mean optical density (arbitrary units) normalized to actin. (B) Control and LyGDI siRNA-transfected samples were tested for FcγR expression by flow cytometery analysis. Overlay histograms show FcγR staining in secondary-only antibody controls (gray), control siRNA-transfected (black) and si-LyGDI-transfected (red). (C) Control and LyGDI siRNA-transfected PBMs were stained with mouse anti-Rac followed by Alexa flour 555 conjugated to goat anti-mouse IgG, F(ab')2 fragment. (D) The fold change in Rac membrane localization in control versus LyGDI siRNA-transfected PBMs was quantified as described in Materials and Methods.