Figure 1

Expression of FoxP3 and Bcl-xL using a 2A gene sequence in primary CD4⁺ T cells. (a) Schematic representation of the retrovirus construct expressing Bcl-xL and FoxP3. ψ, packaging signal; 2A, picornavirus self-cleaving 2A sequence. (b) Naive CD4⁺ T cells from C57BL/6J mice were stimulated with anti-CD3 plus anti-CD28 antibodies. On day 2/3, T cells were transduced with retroviral vectors expressing green fluorescent protein (GFP) (Mig), GFP with FoxP3 (Mig-FoxP3), GFP with Bcl-xL (Mig-Bcl-xL), or GFP with Bcl-xL and FoxP3 (Mig-Bcl-xL-2A-FoxP3). On day 8 of primary culture, GFP⁺ CD4⁺ T cells were sorted, and protein expression of FoxP3, Bcl-xL, and β-actin was determined by Western blotting. Data are representative of three independent experiments. (c) Intracellular FoxP3 expression on day 6 of primary culture was analyzed by flow cytometry after gating on live CD4⁺ T cells. Data are representative of three independent experiments.