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Fig. 6 | Retrovirology

Fig. 6

From: Extracellular vesicles from HTLV-1 infected cells modulate target cells and viral spread

Fig. 6

HTLV-1 100 k EVs enhance viral transmission in dendritic cells. a A diagram representing a timeline of the experimental design and treatment conditions from day 0 to day 9. b Flow cytometry analysis of the EV uptake by mDCs was performed in biological duplicate. Dot plots (upper panels) contain a P1 (red) and P2 (green) gated mDC populations, where the P1 was used to evaluate percent change (% ), increases in size and morphology, of P1 control compared to EV treatment. The control mDC populations in P1 and P2 are 33.7% and 49.9%, respectively. Histograms (lower panel) show increase in fluorescence intensity (right shift) as a function of EV uptake by mDCs. c Recipient mDCs were incubated for 5 days with various populations of HTLV-1 EVs (2 k, 10 k, and 100 k EVs) and CEM EVs (control EVs) at a ratio of 1 cell to 10,000 EVs. This was followed by microscopic analysis. Images are representative of three independent experiments. Following microscopy, recipient cells were then treated with irradiated HUT102 cells (HTLV-1 Donor Cells; 10 Gy) and fresh exo-free media for 4 additional days. RNA was isolated from recipient cell pellets and quantitated by RT-qPCR for viral env RNA. The RNA levels for the EV input are denoted by blue bars (control), while recipient cells treated with the input EVs by the orange bars (Recipient). The recipient mDC viability was analyzed at day 9 (cultured in biological triplicates). Vertical dashed bar (---) separates additional control EVs on the right-hand side for Total EVs (from HUT102) and Control EVs (from CEM). A two-tailed student t-test was used to compare control cells with recipient cells and to evaluate statistical significance with “*” for p-values ≤ 0.05, “**” for p-values ≤ 0.01, and “***” for p-values ≤ 0.001, indicating the level of significance

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