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Mutations of the Rho insert region do not perturb protein stability. Hygromycin-selected NIH 3T3 cells stably expressing HA epitope-tagged Rac1(61L) or the indicated insert mutant protein were lysed. The levels of Rac1 proteins expressed in each lysate were determined by Western blotting using anti-HA MAbs (BAbCO). Comparable levels of expression were detected in stable transfections of NIH 3T3 cells or in transient overexpression analyses in COS-7 cells (data not shown).
