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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 3.

Figure 3

shRNA transduction does not grossly affect differentiation to macrophages derived from FL-CD34+ cells. (A) FL-CD34+ cells (2 × 105) were transduced with lentiviral vectors encoding sh1005 or muCCR5 shRNA as a control. Mock are cells without shRNA transduction. The cells 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 the macrophages. The attached cells were gently scraped off the bottom of the plate and 1 × 105 cells were analysed for CD14, CD11b and CD206 expression by flow cytometry. (B) 1 × 105 macrophage cells were incubated with an MOI of 50 Texas Red-conjugated Zymosan particles for 90 min. Cells were then washed with cold PBS for three times and were analysed by fluorescence microscopy and flow cytometry. The flow cytometry results are exhibited as EGFP versus Zymosan bioparticle (Texas Red) dot plots. (C) Surface CCR5 staining of macrophages derived from sh1005 and muCCR5 shRNA-transduced cells.−, isotype control; +, CCR5 staining