Figure 5.

The C termini of ACAT1-365 and ACAT2-345 are oriented differently in the ER membrane. (A) ACAT1-365 and ACAT2-345 mRNA were translated in vitro in the presence and absence of rough microsomes and glycosylation inhibitor, as described in Figure 2B. The position of the glycosylated ACAT2-345 product (lane 5) is indicated by the arrow. (B) AC29 cells were transiently transfected with ACAT1-365 (a and b) or ACAT2–345 (c and d). After permeabilization with digitonin (a and c) or saponin (b and d), cells were stained with anti-FLAG M2 C-term antibody as described in Figure 2A. (C) ACAT1-365 and ACAT2-345 membrane topology models.