FIG. 4.

Membrane orientation of UL45-2. COS7 cells were transfected with pAC265 (wild-type UL45-2, lanes 1 and 2), pMM336 (R4N mutant, lanes 3 and 4), or pMM337 (E111N mutant, lanes 5 and 6). Cytoplasmic extracts were prepared at 40 h posttransfection, incubated for 2 h at 37°C in the presence (lanes 2, 4, and 6) or absence (lanes 1, 3, and 5) of endo F, electrophoresed on a denaturing gel, transferred to nitrocellulose, and probed with LSU32 and then with ¹²⁵I-protein A. Molecular weights (in thousands [K]) are noted at the right.