Figure 5.

TIEG1 expression restores Foxp3 expression in Itch^−/− T cells. (a) Immunoblot analysis of TIEG1, Smad2 and Smad4 in lysates of Itch^+/+ and Itch^−/− CD4⁺CD25⁻ cells stimulated with anti-CD3 plus anti-CD28 with or without TGF-β (5 ng/ml). (b) Flow cytometry of CD4⁺CD25⁻ cells transduced with pMX or pMX-TIEG1, then stained intracellularly with phycoerythrin-labeled anti-Foxp3 after 6 d. Numbers in quadrants indicate percent Foxp3⁺GFP⁻ cells (top left) or Foxp3⁺GFP⁺ cells (top right). (c) Immunoblot analysis of Foxp3 in lysates of CD4⁺CD25⁻ cells transduced with pMX or pMX-TIEG1; GFP⁺ cells were sorted by flow cytometry and treated with TGF-β. Bottom, same lysates probed with anti-Grb2 (loading control). Data are representative of three independent experiments.