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Fig. 4. Rac1 modulates macropinocytosis of E-cadherin. (A) Cells were serum-starved overnight, stimulated with EGF for 10 minutes and lysed. Then, levels of Rac1-GTP in equivalent concentrations of cell lysate were determined by PAK assay and compared with total Rac1 levels. Change in Rac1-GTP levels was quantified by densitometry. (B) MCF-7 cells were transiently transfected with either (a) wt-Rac1-GFP, (b) CA-Rac1-GFP or (c) CA-Cdc42-GFP (all in green), fixed and stained for endogenous E-cadherin (red) and nuclei (DAPI, blue) before confocal microscopy analysis. Note localization of E-cadherin to enlarged endosomal compartments labeled for CA-Rac1-GFP in b (arrows). Second, third and fourth panels are magnifications of boxed areas shown in respective first panels. Bars, 20 µm for all first panels; 2 µm for all other panels.