Fig. 3

Incorporation of PR-defective Gag-Pol into wt virus particles. a Schematic representations of HIV-1 Gag and Gag-Pol expression vectors. Indicated are the HIV-1 Gag protein domains MA (matrix), CA (capsid), p2, NC (nucleocapsid), p1 and p6, and pol-encoded p6*, PR, RT and IN, all as described in the Fig. 1 caption. “X” denotes a substitution mutation at the PR catalytic Asp residue. The C-termini of constructs fsDmyc and CSMfsDmyc are attached with myc epitopes. b-e 293T cells were transfected with 10 µg of wt plasmid DNA plus the indicated amounts of fsDmyc or CSMfsDmyc plasmid DNA. At 48 h post-transfection, cells and supernatants were collected, prepared, and subjected to Western immunoblotting. Gag proteins were probed with anti-p24CA and anti-p17MA monoclonal antibodies. Panel c was derived from panel b following sequential probing with anti-RT, anti-Myc or anti-IN antibodies. Shown is a representative from three independent experiments