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Fig. 7 | Retrovirology

Fig. 7

From: Fused in sarcoma silences HIV gene transcription and maintains viral latency through suppressing AFF4 gene activation

Fig. 7

Knockdown of FUS expression enhances reactivation of HIV latency by JQ1. a Depletion of FUS delays the establishment of HIV latency. J-LTR Luc (red line) or J-LTR-Luc FUS KO cells (green line) were transduced with pseudotyped HIV-pNL4-GFP virus, where a GFP reporter in inserted instead of the env gene. Cells were sorted based on their GFP expression (day 0 post infection) and further grown for the indicated time days post infection to allow them to gradually enter viral latency. GFP expression was monitored at the indicated time points by FACS analysis as a reference for entry into viral latency. b Reactivation of latent cells. At 60 d.p.i., transduced J-LTR Luc or J-LTR-Luc FUS KO cells were sorted based on their GFP expression for GFP(−) cells. Cells were then treated for 24 h. with either PMA or JQ1 activators, at the indicated concentrations, and subjected to FACS analysis to monitor their GFP expression, which corresponds to viral reactivation. Error bars indicate mean ± SD from triplicates. c Knockdown of FUS expression enhances reactivation of HIV latency by JQ1. 2D10 latent cells were introduced with either FUS-specific or scrambled (control) siRNA oligos. 72-h post transfection, cells were treated with JQ1 in the indicated concentrations. 24 h post treatment HIV gene expression was analyzed by FACS, monitoring d2GFP. d HIV RNA levels are elevated upon FUS KD in 2D10 cells and treatment with JQ1. Control 2D10 latent cells (harboring scrambled siRNA), or cells where FUS expression was depleted by specific siRNA were treated with 0.2 μM JQ1 and subjected to RT-qPCR with specific primers amplifying the indicated HIV gene targets - Gag and Rev. Results are presented relative to GAPDH mRNA levels. Also presented is a western blot confirming KD of FUS expression

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