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Fig. 6. Effect of a GFP-ß2-chimaerin GAP-defective mutant on CXCL12/PMA-induced adhesion. (A) Vectors encoding GFP, GFP-ß2-chimaerin or a ß2-chimaerin GAP-defective mutant bearing a deletion of the EIE sequence (GFP-{Delta}EIE) were transfected into HEK293 cells. At 24 hours post-transfection, cells were lysed and Rac-GTP levels determined by pull-down assays with PAK-RBD-GST. One-tenth of each sample was analyzed for total Rac level and ß2-chimaerin expression. (B) Jurkat T cells were transfected with the indicated plasmids; cells were sorted 24 hours post-transfection and GFP-positive cells tested for adhesion as in Fig. 5. Data represent the mean ± s.d. of at least three independent experiments performed in triplicate.





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