HIV-1 integration and infection of PB-CD34 cells using a dual-reporter pseudovirus. Pseudoviruses constructed with RGH backbone and envelope from HxB2 (X4-tropic) and JR-FL (R5-tropic). (A) Schematic representation for active infection indicated by eGFP fluorescence (green), integrated viral DNA (latency) by mCherry fluorescence (red), and active replication from integrated genome (yellow; arrows in panels B-C). Confocal microscopy was used to examine eGFP and mCherry fluorescence signals 2 days after infection. Representative images of cells showing single or dual mCherry/eGFP signals in Jurkat cells (B) or mock or HCMV-infected PB-CD34 cells (C) at day 2 after RGH/HxB2 pseudovirus infection, with cell count data shown from 4 independent PB-CD34 experiments. Columns represent data as mean ± SEM. Average number of cells with integrated genome is shown below. Original magnification ×400 for panels B-C. *P < .05, **P < .01.