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. 2013 Jan 29;21(4):786–795. doi: 10.1038/mt.2012.284

Figure 2.

Figure 2

Establishment of JLTRG-R5 T-cell line derived cells expressing anti-HIV genes. (a) JLTRG-R5 cells were transfected with the rhesus or human-rhesus hybrid CCR5-TRIM5α targeting vector and CCR5 ZFNs, and were analyzed for targeting by Southern blot. (b) Protein expression of restriction factors targeted to CCR5, probed by α-HA (TRIM5α), α-myc (APOBEC3G D128K) and α-Rev (Rev M10). The Coomassie stained blot is a loading control. (c) Quantification of bands from western blots in panel b using ImageJ software and normalized to hTRIM, rev-APO and rev lanes, respectively. Error bars represent the variation between quantification from two gels. (d) FACS plots showing ZFN-mediated disruption of the untargeted CCR5 allele in CCR5-IRES-puro cells. See also Supplementary Figure S5. a, APOBEC3G D128K-myc; FACS, fluorescence-activated cell sorting; h, human TRIM5α HA; hrh, human-rhesus hybrid TRIM5α-HA; r, Rev M10; rh, rhesus TRIM5α-HA; WT, wild type; ZFN, zinc finger nuclease.