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Fig. 3 | Retrovirology

Fig. 3

From: HIV-1 latency and virus production from unintegrated genomes following direct infection of resting CD4 T cells

Fig. 3

Kinetics of latency reversal and virus production from sorted GFP-negative cells following in vitro infection of resting CD4 T cells from 3 donors. The 14-day latency protocol from Fig. 2a was followed using cells from 3 HIV-negative donors following infection with equal amounts of HIV-1 in order to investigate kinetics of latent virus activation and donor variability. For added precaution against de novo infection, Indinavir was added at day 0 and 7 p.i., while efavirenz was added at day 5 p.i. for donors 2 and 3. a Percent GFP+ cells after Pro/TSA or DMSO carrier control treatment of sorted GFP-negative cells. Percent GFP+ cells prior to sort (No RAL, +RAL): Donor 1: 4.9 %, 3.3 %, Donor 2: 16.9 %, 22.4 %, Donor 3: 21.4 %, 17.3 %. b Virions released into culture medium measured by RT-qPCR for genomic viral RNA. Lower limit of quantitation was 1 virion/µl. c The difference in virus output from the No RAL and +RAL cultures decreases over time following latency reversal. Graphs present the ratio of No RAL virus output vs. +RAL virus output over the indicated interval. Output over each interval for No RAL and +RAL cells was calculated by subtracting the virions/µL at the earlier indicated time from the later time, then the ratio of No RAL/+RAL was graphed. For example, for Donor 1 during the first 14 h (“0–14”) the No RAL cells released 7.3 times as many virions as the +RAL cultures but over the 41–65 h interval, the No RAL cells release 2.2 times as many virions as the +RAL cells. Red line indicates equal output from the No RAL and +RAL cells. d The difference in virus output per GFP+ cell from the No RAL and +RAL cultures decreases over time following latency reversal. The ratio of No RAL virus output per GFP+ cell vs. +RAL virus output per GFP+ cell over the indicated interval. Output during each interval, calculated as in 3C was then divided by the percent GFP+ cells at the end of each interval. For example, for Donor 1 during the first 14 h (0–14) each GFP cell in the No RAL infections released 9.5 times as many virions as the +RAL GFP+ cells but during 41–65 h interval, the No RAL cells released 2.9 times as many virions per GFP+ cell. Red line indicates equal output from the No RAL and +RAL cells

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