Figure 8.
(A) In vitro budding reactions with microsomes prepared from cells expressing both 3HA-Emp46p and 2myc-Emp47p. Ten percent of the total reaction (T) and budded COPII vesicles isolated after incubation with (+) or without (−) COPII proteins was analyzed by SDS-PAGE followed by immunoblotting with an anti-HA or anti-myc. Sec61p as negative control and Sec22p as positive control were detected with polyclonal antibodies. (B) In vitro budding reactions with microsomes prepared from strains expressing C-terminal mutants of Emp46p. The same protocol as in A was used except that the reaction time was 15 min.