Fig 7.

Estimate of the abundance of endogenous palmitoylated bCdc42 in neonatal brain. (A) Postnuclear lysates of mouse neonatal brain and adult kidney were fractionated by ultracentrifugation. The pellet was subjected to acyl-RAC as described in Materials and Methods. Acyl-RAC samples and input (10% of total) were subjected to immunoblotting using bCdc42-specific and Cdc42 antibodies. Cdc42 antibody detects both Cdc42 and bCdc42 isoforms. Band intensity was quantified. (B) Postnuclear lysates from neonatal brain and adult kidney were separated by ultracentrifugation into supernatant (S) and pellet (P) fractions. Proteins were extracted from the pellet fraction and mixed with GST-RhoGDIα bound to glutathione-Sepharose. GST-RhoGDIα unbound (Un) and bound (B) fractions were collected. bCdc42 and Cdc42 were detected by immunoblotting. Band intensity was quantified, and the relative proportions in each fraction are shown.