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Fig. 4. Cell surface expression and localization in plasma-membrane protrusions of prominin-1.s3 and s6 splice variants in CHO cells. (A,B) CHO cells stably transfected with prominin-1 splice variants s1 (positive control), s3, s4, s5, s6 or with vector DNA without insert (MOCK) were either double cell-surface labeled with the mAb 13A4 and FITC-conjugated wheat-germ agglutinin (WGA) (green) followed by Cy3-conjugated anti-rat secondary antibody (red) (13A4) (A) or labeled after paraformaldehyde fixation and Triton-X-100 permeabilization with mAb 13A4 followed by Cy3-conjugated anti-rat secondary antibody (red) (B). (A) Single optical sections at the level of the middle of the cells. (B) Composite pictures of eight optical sections. Scale bars, 45 µm (A), 60 µm (B). (C) Ultrathin cryosections of CHO cells transfected with s3 and s6 splice variants were stained with either the mAb 13A4 (s3) or E3 antiserum (s6) followed by rabbit anti-rat IgG/IgM and/or 15-nm Protein-A/gold. Scale bars, 245 nm (s3); 420 nm (s6).

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