Figure 2.

p200 associates with the Golgi apparatus. (A) Production of full-length HA-p200 in 293 HEK-EBNA cells. Detergent extract (60 μg) from stably transfected cells were analyzed by Western blotting to detect HA-tagged p200 (arrowhead). (B) Indirect immunofluorescence. BHK21 cells were transiently transfected with the construct encoding HA-tagged protein by using Lipofectamine and were processed for immunofluorescence by using antibodies against the HA epitope (12CA5) and either mannosidase II or ERGIC53/58, as indicated. Each horizontal pair of panels corresponds to a field of view seen with different filters. The distribution of HA-p200 and mannosidase II in cells exposed to 3.5 μM BFA for 10 min before fixation is shown in the middle panels.