(A) Jurkat T cells were transduced with GFP or YFP-2A-FOXP3 lentiviral (LV) vectors; stable expression was monitored over time by GFP/YFP fluorescence. (B) FOXP3 and SAP expression was measured in LV-transduced Jurkat T cell lysates by Western blot. Numbers represent quantitation of SAP expression by spot densitometry, normalized to LV-GFP transduced cells. (C) LV-transduced Jurkat T cells were lysed for chIP using antibodies specific for FOXP3, histone H3, or an isotype IgG control. Immunoprecipitated DNA was then subjected to PCR using primers specific for the SH2D1A promoter, or the RPL30 housekeeping gene. Results are representative of 3 independent experiments. (D) ChIP analysis was performed on activated human Tregs and Tcons using FOXP3 or IgG isotype antibodies. Immunoprecipitated DNA was then subjected to PCR using primers specific for the SH2D1A promoter, or the RPL30 housekeeping gene. Data are representative of 2 independent experiments.