The ability of hA3G to bind Gag is insufficient for its incorporation into HIV-1. 293T cells were co-transfected with plasmids coding for hGag and either hA3G Y124A (A) and W127A (B). Left panel: Western blot of cell lysates were probed with either anti-HA (top) or anti-β-actin (bottom). Middle panel: Western blot of virion lysates were probed with either anti-HA (top) or anti-p24 (bottom). Right panel: Western blots of rabbit anti-p24 immunoprecipitates were probed with either anti-HA (top) or mouse anti-p24 (bottom). C. Relative amounts of hA3G Y124A in the LMM, RA LMM and HMM hA3G complex were determined by Western blot and graphically presented. The bar graphs represent the means of results of experiments performed at least three times, and the error bars represent standard deviations.