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. 2014 Oct 31;9(10):e111437. doi: 10.1371/journal.pone.0111437

Figure 2. Phenotypic selection using SBP-ΔLNGFR.

Figure 2

293Ts were transiently transfected or lentivirally transduced with pHRSIN-SE-PGK-SBP-ΔLNGFR-W (encoding EGFP and SBP-ΔLNGFR; A) or pHRSIREN/β2 m-PGK-SBP-ΔLNGFR-W (encoding shRNA to β2 m and SBP-ΔLNGFR; B) and stained with streptavidin-APC plus/minus anti-HLA-A2-PE. Transfected/transduced cells are either GFP+/streptavidin-APC+ or HLA-A2-low/streptavidin-APC+ (dashed circles). Primary human CD4+ T cells were lentivirally transduced with the same constructs then selected using Dynabeads Biotin Binder. Purity of cells before (green or blue) and after (red) selection was assessed by GFP fluorescence (C) or staining with anti-HLA-A2-PE (D). Transduced cells are either GFP+ or HLA-A2-low (dashed boxes). Background staining of untransfected/unstransduced controls is shown (grey).