FIG. 7.
Low levels of HIV-1 structural and Nef protein expression compared to Rev protein expression by astrocytes. Immunoblotting or immunoprecipitation of viral antigens expressed in U251MG (lanes 1 and 2) and HeLa (lanes 3 and 4) cells after cotransfection with the HIV-1 proviral plasmid pNL4-3 and pCMV-hGH (lanes 2 and 4) or pCMV-hGH transfection efficiency reporter alone (lanes 1 and 3). Cell lysates were normalized for hGH production, and viral proteins were detected with polyclonal HIV-immune serum (BB10; 1:5,000) (A) or anti-Nef15–27 (1:500) (B). (C) Rev protein detected by immunoprecipitation with sheep antiserum to HIV-1 Rev (1:500) from HeLa and U251MG cells labelled with [35S]methionine-cysteine for 16 h, 48 and 72 h after transfection, respectively.