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. Author manuscript; available in PMC: 2015 Feb 25.
Published in final edited form as: J Gene Med. 2010 Mar;12(3):255–265. doi: 10.1002/jgm.1440

Figure 9.

Figure 9

Stable knockdown of CCR5 expression on macrophages derived from FL-CD34+ cells rendered resistance to R5 tropic HIV-1 NFN-SX infection. FL-CD34+ cells (2 × 105) were transduced with lentiviral vectors expressing sh1005 or muCCR5 shRNA. The cells were harvested 3 days after virus transduction, sorted by the EGFP expression. FL-CD34+ cells (1 × 105) stably expressed sh1005 or the muCCR5 shRNA were treated with 20 ng/ml of IL-3, 50 ng/ml of SCF and 20 ng/ml of MCSF for 9 days and then with 5 ng/ml of GM-CSF for 5 days to differentiate to macrophages. The cells were then infected with either wild-type NFN-SX or VSV-G pseudotyped NFN-SXΔEnv, and cell free culture supernatant (1.0 ml) was harvested every 7 days post-infection for 21 days. Virus replication was monitored by measuring p24 antigen in culture supernatants. Mock, cells not infected by NFN-SX; −, nontransduced cells