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Fig. 5. Kinetics of activation of Cdc42 and Rac upon infection. HeLa cells in suspension were centrifuged for 20 seconds in DMEM alone (control lane) or with bacteria (other lanes), and incubated for the indicated times at 37°C. Cells were lysed and incubated with GST-CRIB. GST-CRIB-associated proteins were pulled-down using glutathione-Sepharose and analysed by western blotting using anti-Rac1 (A) and anti-Cdc42 (C) antibodies. Aliquots of total cell lysates were immunoblotted for total Rac1 (B) and total Cdc42 (D), showing that the total amount of each small GTPase was identical at all time points. Data are representative for four experiments.
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