Fig. 2.

Removal of MA ameliorates NC15A assembly and processing defects. (A) 293T cells were transfected with indicated constructs. At 48 h post-transfection, cells and culture supernatants were collected, prepared, and subjected to Western immunoblotting probed with an anti-p24CA antibody. (B) Gag proteins from medium or cell samples were quantified by scanning mutant and wt p24gag-associated band densities from immunoblots. Ratios of total Gag protein levels in the media to those in cells were determined for each construct and compared with wt release levels by dividing the release ratio for each mutant by the wt ratio in parallel experiments. Error bars indicate standard deviation. ^∗p < 0.05. (C) 293T cells were transfected with indicated constructs. At 48–72 h post-transfection, culture supernatants and cells were collected and subjected to Western immunoblotting. (D) Total Gag proteins (wt or mutant Gag precursor) from medium or cell samples were detected with an anti-p24CA monoclonal antibody and quantified by scanning wt and mutant Gag precursor band densities from immunoblots. Ratios of Gag in media to Gag in cells were determined for each construct, and compared with wt release levels as described above. Error bands indicate standard deviation. ^∗p < 0.05.