Subcellular localization of CD33 and His-M195FANCF.

HeLa cells were transiently transfected with pcDNA3-CD33 and plated on glass coverslips. Either intact (A,B) or detergent-permeabilized cells (C-F) were then incubated with His-M195FANCF, and the localization of CD33 and His-M195FANCF was assessed by fluorescence microscopy. Similar results were also obtained with transfected 293 cells (data not shown). Antibodies used were anti-CD33 monoclonal antibody followed by Texas red–conjugated goat anti–mouse IgG (A,C,E,F) and anti-FANCF polyclonal antibody followed by FITC-conjugated goat anti–rabbit IgG (B,D,G). (F) Double exposure for CD33 expression and for visualization of blue-stained nuclei by Hoechst dye.