FIGURE 15.

Cell localization of Oatp1d1 after mutations of conserved N-glycosylation sites. Mutagenesis of N122Q, N133Q, N499Q, and N512Q, as well as simultaneous mutation of three (N122/133/499Q) and four asparagine residues (N122/133/499/512Q), respectively, was performed, and images were obtained with immunofluorescence analysis by confocal microscopy. Immunocytochemistry was performed with FITC, which binds to the primary Xpress antibody and stains the protein in green. Nuclei are dyed in blue with DAPI, and plasma membranes are stained in red after binding of primary antibody Na,K-ATPase and Cy3-conjugated IgG secondary antibody (all anti-mouse). White arrows indicate examples where protein is present in the cell membrane. The color turns to orange because of the overlap of green and red, or it remains light green when overexpression of protein is dominant over red-stained membranes. Cytosolic forms are seen as green areas in the cytoplasm.