FIG 5.

The single-positive protein population is not the result of new rounds of HIV-1 infection. (A) hTNF-α was used at a final concentration of 10 ng/ml to stimulate J89 cells for 24 h simultaneously with 20 μg/ml AMD3100 or 100 μg/ml AMD3100 or without AMD3100 (negative control). A PrimeFlow RNA assay was performed to distinguish the double-negative population, the viral mRNA-single-positive population, the viral mRNA/protein-double-positive population, and the protein-single-positive population. No blocking of the p24 protein-single-positive population was observed in the presence of the CXCR4 inhibitor AMD3100. (B) J-Lat latency reactivation. RMD at a final concentration of 5 nM and hTNF-α at a final concentration of 10 ng/ml were used to stimulate 2.5 × 10⁶ J-Lat cells at two time points, 24 h and 48 h. Unstimulated cells were used as a control. A PrimeFlow RNA assay was performed as described in the text to distinguish four populations: the double-negative population, the viral mRNA-single-positive population, the viral mRNA/protein-double-positive population, and the protein-single-positive population.