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. 2007 Mar 7;104(11):4571–4576. doi: 10.1073/pnas.0700298104

Fig. 5.

Fig. 5.

T cell stimulation antagonizes FOXP3 recruiting HDAC9. (A) HA-FOXP3a transfected Jurkat E6.1 T cells (10 × 106) were not stimulated, or stimulated with plate-bound TCR Vβ 8.1 plus soluble anti-CD28 for 4 h (with or without 400 nM TSA, indicated above lanes), lysed, and equal nuclear extracts were immunoprecipitated with anti-HA probe (F-7), then analyzed by immunoblotting with anti-HDAC9 H-45 (lanes 3–6). The input nuclear extracts of TSA treated cells, with or without stimulation, were also immunoblotted with anti-HDAC9 H-45 (lanes 1 and 2). (B) In vitro activated and expanded human CD4+CD25+ T cells were treated with or without 400 nM TSA and lysed, and equal nuclear extracts were immunoprecipitated with anti-FOXP3 mAb 221D or control IgG, then analyzed by immunoblotting with anti-HDAC9 H-45 (Right, lanes 3, 4, and 5). The input nuclear extracts of TSA treated or untreated cells were also immunoblotted with anti-HDAC9 H-45 (Left, lanes 1 and 2).