FIG. 2.
Env ERVWE1 expression in transfected human cells. (A) For Western blot analysis (WB), cells transfected with wild-type HERV-W (WT) and antisense orientation (control) plasmids were separated by SDS-10% PAGE. The blots were probed with anti-SU-EnvW and anti-TM-EnvW. For immunoprecipitation analysis (IP), cells expressing the WT envelope protein were immunoprecipitated with the anti-TM MAb and the anti-p24 MAb (control). Immunoprecipitated proteins were analyzed by SDS-10% PAGE. Arrowheads indicate the positions of gPr73, gp50 SU, and gp24. The asterisk indicates the position of the glycosylated SU protein. (B) Furin cleavage site mutant analysis. Cell lysates expressing WT and furin cleavage site mutant (AAAR) envelope proteins were separated by SDS-10% PAGE and probed with anti-TM-EnvW. (C) Furin cleavage site mutant cell surface expression. Cells expressing the WT (RNKR, shaded area), furin cleavage site mutant (AAAR, shaded area), or antisense orientation (control, white area) envelope were stained with the anti-TM MAb. The cells were analyzed by flow cytometry.