Characterization of the association between Gag and HIV-1 RNA. (A) Measurement of the association between Gag and HIV-1 genomic RNA. HIV-1 genomic RNA immunoprecipitated with the Gag was characterized by qPCR using a primer/probe set targeting the unspliced RNA transcript. (B) Measurement of the association between Gag and spliced HIV-1 mRNA. The same cDNA preparation described above was subjected to qPCR using a primer/probe set specific for env mRNA and rev mRNA sequences. The copy number in each sample was adjusted for input by the cell number and for transfection efficiency by GFP expression from a co-transfected reporter construct. The amount of NL4-3 RNA was set at 100%. Means and SD of three independent experiments are shown. *, indicates p < 10-4 and significant deviation from the wild-type copy number as determined by Student's t test.