FIG. 8.
Mid2 and Hcs77 are O-mannosylated. (A) Extracts (50 μg of protein) of yeast strain 1788 expressing Hcs77 (lane 1), Hcs77-HA (lane 2), Hcs77N4Q-HA (lane 3), Hcs77N65Q-HA (lane 4), or Hcs77N4Q,N65Q-HA (lane 5) from YEp352 were subjected to SDS-PAGE followed by immunoblot analysis. (B) Mid2-HA (lanes 1 and 2) and Hcs77-HA (lanes 3 and 4) were immunoprecipitated from extracts (100 μg of protein) of yeast strain 1788. Immunoprecipitates were either treated with α-mannosidase at 37°C for 3 h (lanes 2 and 4) or mock treated with buffer only under the same conditions (lanes 1 and 3). Immunoprecipitates were subjected to SDS-PAGE and immunoblot analysis. Sizes are indicated in kilodaltons. IgG, immunoglobulin G.