FIG. 5.

Env incorporation of NL(AD8)-NX viruses carrying gp41 CTs and matrix proteins derived from HIV-1 present at the early or late stages in patient 153. Virus stocks of the indicated molecular clones were obtained by transfecting 293T (A) and HeLa cells (B) or by infecting MT4.R5 cells (C), PBMC (D), or MDMs (E), as described in Materials and Methods. Env incorporation into virions was assessed, using viruses purified by centrifugation through a 20% sucrose cushion. Viral pellets were lysed in TNE buffer containing 1% Triton X-100 and gp120 was quantified by ELISA. The results shown were obtained from a representative experiment performed in duplicate. Errors bars indicate standard deviations. (F) Western blot analysis of Env incorporation into virions produced by HeLa cells. The upper panel shows the blot probed with anti-gp120 and anti-p24 antibodies. The positions of the Env glycoprotein gp120, the Gag precursor Pr55, the Gag processing intermediate Pr41 and the processed p24 capsid protein are indicated. The blot was stripped and reprobed with an anti-gp41 antibody to detect the Env glycoprotein gp41 (lower panel; see Materials and Methods).