Quantitation of viral RNAs in LNMCs draining immunization sites. Total RNA extracted from LNMCs draining either SIV gag or RhCMV pp65 immunization sites, one animal per time point, were subject to real-time PCR to quantitate SIV doubly spliced (tat) (A), singly spliced (vif) (B), and unspliced (gag) (C) RNA. LNMCs draining the RhCMV pp65 immunization site (Gray). LNMCs draining the SIV gag immunization site (Black). Copy numbers of SIV RNAs were normalized to macaque GAPDH mRNA, and the results for each time point following immunization (PI, post immunization) are shown as fold change relative to RNA analyzed on day 60-post immunization from the same animal, as no differences between LNs draining gag or pp65 immunizations were found at this time point. Error bars are the standard error of the mean of replicate analyses. Statistical analysis compared viral RNA between LNMCs draining SIV and CMV immunization sites in the same animal on the same day. Most measurements were repeated in at least two separate experiments with identical results.