FIG. 1.

Mouse CD82-mRFP1 matures normally and demonstrates intracellular distribution similar to that of human CD82. RAW 264.7 cells were transduced to stably express CD82-mRFP1. The cells were pulse-labeled for 15 min with [³⁵S]cysteine/methionine and chased for 0, 90, or 180 min (A) or labeled for 3 h (B). The cells were lysed in 1% NP-40, and proteins were immunoprecipitated with anti-mRFP1 antisera. At each time point, half of the sample was treated with endoglycosidase H (A) or F (B) for 1 h at 37°C. Samples were run on a 10% SDS-polyacrylamide gel, and polypeptides were visualized by autoradiography. The “CHO” labels indicate the presence of N-linked glycans, and complex-type sugars (C.T.) were present. Molecular sizes (in kilodaltons) are given at right. (C) HeLa cells expressing mouse CD82-mRFP1 were fixed, permeabilized, and stained with an anti-human CD82 antibody. Shown is an image of a single cell illustrating mouse CD82-mRFP1 (left, red), human CD82 (center, green), and a merged image (right). Bar, 5 μm.