Subcellular distribution and prenylation of mutant and wild-type Rac2 proteins.

(A) Transduced and sorted NIH3T3 cells expressing EGFP alone, Rac2^D57NΔCT, Rac2^D57NΔM, and wt Rac2 were fractionated into cytosol (S100) and crude membrane (P100) fractions. Equivalent proportions of each fraction were resolved by SDS-PAGE followed by Western blotting using anti-Flag (M2) antibody to detect various Rac2 proteins. S = S100; P = P100. (B) Transduced and sorted NIH3T3 cells expressing EGFP alone, Rac2^D57NΔCT, Rac2^D57NΔM, and wt Rac2 were labeled with [³H] mevalonolactone, and mutant and wt Rac2 proteins contained in these cells were immunoprecipitated with anti-Flag (M2) antibody and analyzed by SDS-PAGE. The gel was treated with EN³HANCE and exposed to Kodak XOMAT film.