FIGURE 2.

CRMP2-K374A mutant expresses robustly in CAD cells. A, immunoblot with anti-FLAG antibody from CAD lysates transfected with wild type (CRMP2) or the putative single site SUMOylation conjugation site mutant (CRMP2-K374A) showing comparable levels of expression of the proteins. Representative monochrome images of wild type CRMP2 (B) or mutant CRMP2–374A (C) expressed in CAD cells. Forty eight hours after transfection, the cells were permeabilized and then labeled with a mouse monoclonal antiserum directed against the FLAG epitope and visualized by incubation with fluorescent-conjugated Alexa 488 goat anti-mouse secondary antibody. Both proteins showed similar patterns of expression throughout the cytoplasm of the transfected cells. Scale bar, 20 μm.