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Fig. 2. Trans-endocytosed CD47 is sorted into multivesicular bodies. (A,B) CHO–SHPS-1 (A) or CHO-Ras (CHO) cells (B) were cocultured with CHO-CD47 cells for 1 hour. The cells were then fixed, and stained with a mAb to CD47 and immunogold-labeled secondary antibodies. Ultrathin sections were prepared after silver enhancement and stained with uranyl acetate and lead citrate as described in Materials and Methods. MVBs in CHO–SHPS-1 (A) and CHO-Ras cells (B) are shown. Bar, 100 nm. (C,D) CHO–SHPS-1 cells were cocultured with CHO-CD47 cells for 1 hour. The cells were then fixed and subjected to double-immunostaining with a mAb to CD47 (green) and pAbs to cathepsin D (red) in the presence of 0.1% Triton X-100. A higher magnification image of boxed region in C was shown in D. The dotted line in C indicates a CHO–SHPS-1 cell. Bar, 10 µm.
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