FIG. 3.

Western blot analysis of havcr-1 mutants expressed in dog cells. Cytoplasmic extracts of flag (lanes 1 and 2), d1− (lanes 3 and 4), mut1 (lanes 5 and 6), mut2 (lanes 7 and 8), and mut3 (lanes 9 and 10) cells or of control DR2 cells (lanes 11 and 12) were prepared in RSB–1% NP-40. Cell extracts containing 20 to 25 μg of total protein were treated with 500 U of PNGase F (+ [lanes 2, 4, 6, 8, 10, and 12]) or mock treated (− [lanes 1, 3, 5, 7, 9, and 11]), separated by SDS–12.5% polyacrylamide gel electrophoresis, transferred to PVDF membranes, and probed with rabbit anti-GST2 Ab directed against the TSP-rich region of havcr-1. The asterisk marks the 67-kDa band corresponding to a form of havcr-1 containing three N-glycans. Arrowheads point to 64-kDa havcr-1 forms containing two N-glycans. Arrows point to 62-kDa havcr-1 forms containing only one N-glycan. The positions of prestained molecular mass markers and their sizes are shown on the right.