PMC:1447668 / 31566-32998 JSONTXT

{"project":"sentences","denotations":[{"id":"T173","span":{"begin":0,"end":95},"obj":"Sentence"},{"id":"T174","span":{"begin":96,"end":392},"obj":"Sentence"},{"id":"T175","span":{"begin":393,"end":472},"obj":"Sentence"},{"id":"T176","span":{"begin":473,"end":592},"obj":"Sentence"},{"id":"T177","span":{"begin":593,"end":901},"obj":"Sentence"},{"id":"T178","span":{"begin":902,"end":985},"obj":"Sentence"},{"id":"T179","span":{"begin":986,"end":1077},"obj":"Sentence"},{"id":"T180","span":{"begin":1078,"end":1282},"obj":"Sentence"},{"id":"T181","span":{"begin":1283,"end":1430},"obj":"Sentence"}],"namespaces":[{"prefix":"_base","uri":"http://pubannotation.org/ontology/tao.owl#"}],"text":"Figure 7 Foxp3 Antagonizes CREB Activation by Blocking Recruitment of Coactivator Protein p300\n(A and B) HEK 293T cells (5 × 105) were seeded into six-well plates 1 d prior to transfection with a CREB or an HTLV-I LTR luciferase reporter vector (200 ng) in the presence or absence of a Foxp3, DFKH, or control expression vector (500 ng) and internal reference plasmid (pGL4-TKhRluc2; 50 ng). Forskolin (10 μM) was added to the appropriate reactions 20 h posttransfection. Reactions were assayed 24 h posttransfection for luciferase activity (A) and CREB-1 and ATF-2 DNA-binding activity (B).\n(C) HEK 293T cells (5 × 105) were seeded into six-well plates 1 d prior to transfection with a Gal4 luciferase reporter vector (200 ng) in the presence or absence of Gal4-BD-CREB-1 (500 ng) along with a control, Foxp3, or p300 expression vector (500 ng) and internal reference plasmid (pGL4-TKhRluc2; 50 ng). Cells were harvested at 24 h posttransfection and analyzed for luciferase activity. Relative luciferase activity in (A and C) was normalized to the internal reference control.\n(D) Whole-cell extracts from HEK 293T cells transfected with a control (EGFP) or Foxp3 expression vector together with a p300-HA expression vector were immunoprecipitated with anti-HA monoclonal antibody. Proteins were then separated by SDS-PAGE and immunoblotted with anti-Foxp3 (ab10563) to detect immunoprecipitates and with anti-HA for the lysates. "}