The correlation between the cellular distribution and viral incorporation of hA3G. A.

<p>Wild type and mutant hA3G. The filled rectangles represent the two zinc coordination units. The numbers represent the amino acid positions. <b>B.</b> 293T cells were co-transfected with hGag and either wild-type or mutated forms of hA3G, and the S1 fractions of the cell lysates were subjected to ultra-centrifugation and octyl glucoside treatment, resulting in the LMM, RA LMM and HMM forms of hA3G. The amounts of hA3G in the three forms were determined by Western blot. <b>C.</b> The cellular distributions of wild type and mutant hA3G are graphically shown. <b>D.</b> Western blot of cell lysates were probed with anti-HA (top) and anti-β-action (bottom). <b>E.</b> Western blot of virus like particle lysates probed with anti-HA (top) and anti-p24 (bottom). <b>F.</b> The relative amounts of mutated hA3G in the cell or viral lysates were normalized to wild-type hA3G, and then a ratio of viral to cellular hA3G was determined and used to measure its ability to be packaged into virions. The bar graphs in panel C and F represent the means of results of experiments performed at least three times, and the error bars represent standard deviations.