FIGURE 2.

Immunofluorescence microscopy analysis of the expression of the RhBG C terminus mutants in HEK293 cells. HEK293 clones transfected with an empty pcDNA3 vector (Empty), or expressing wild-type (RhBG), or mutant RhBG were cultured on poly-l-lysine coverslips and fixed in 4% paraformaldehyde. A, cells were permeabilized in 1% SDS before staining with rabbit anti-RhBG-Cter and Alexa Fluor 488 goat anti-rabbit IgG. B, cells were directly labeled with murine ascites A-08 and Alexa Fluor 488 goat anti-mouse IgG. S422D, S426A, S426D, Y429A, Y429D, T456A, and T456D (not shown) exhibited similar plasma membrane and internal staining in A, and exclusive surface labeling in B. Scale bar, 15 μm.