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Fig. 6 | Retrovirology

Fig. 6

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

Fig. 6

A single round of maximum T cell activation does not induce the expression of all latent unintegrated or integrated viruses. a Experimental design. eFluorhiGFP-negative cells were sorted on day 14 p.i., then cells were stimulated with Pro/TSA, αCD3/CD28 beads or medium only control (Ø). Three days later (day 17) all GFP-negative cells were purified in a second sort and 30,000 cells per well were (re)stimulated in triplicate. Efavirenz was added on days 14 and 17 to sorted cells as an added precaution against virus spread. b Flow cytometric analysis of cells at the end of the first three day treatment. GFP+ cells (light grey dots) are overlaid on GFP-negative cells (black dots). Undivided cells lie above the red lines. c The percentage of GFP+ cells generated after the first round of stimulation calculated based on the total number of GFP+ of the cells placed in the wells on day 14. d Flow analysis of cells after the restimulation. Cells that remained GFP-negative after the first round of stimulation were sorted and equal numbers of these cells were re-stimulated with Pro/TSA, αCD3/CD28beads or medium alone (Ø). e The percentage of GFP+ cells generated after the second round of stimulation calculated based on the total number of GFP+ of the cells placed in the wells on day 17. Mean and SD for each condition are shown

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