Fig. 1.

CD63 fusion proteins mature normally. (A and B) RAW 264.7 cells were transduced to stably express mouse CD63-mRFP1 or HA-CD63. Cells were pulse-labeled for 15 min with [³⁵S]cysteine and methionine and chased for 0, 90, or 180 min (A) or labeled for 3 h (B). Cells were lysed in 1% Brij58 and proteins immunoprecipitated with anti-mRFP1 or anti-HA antibody. 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 12% SDS-polyacrylamide gel and polypeptides visualized by autoradiography. The “CHO” labels indicate the number of N-linked glycans and complex-type sugars (C.T.) present. (C) Human astrocytoma U373 cells expressing mouse CD63-mRFP1 were fixed, permeabilized, and stained with anti-human CD63 antibody. Localization of mouse CD63-mRFP1 (Left) and human CD63 (Center) and the merged image (Right).