Purine-box regulator in T-cell nuclear extracts contains ARRE–DNA-binding subunits Ku80, Ku70 and NF90

Abstract

<p><b>Copyright information:</b></p><p>Taken from "Dynamic binding of Ku80, Ku70 and NF90 to the IL-2 promoter in activated T-cells"</p><p></p><p>Nucleic Acids Research 2007;35(7):2302-2310.</p><p>Published online 27 Mar 2007</p><p>PMCID:PMC1874627.</p><p>© 2007 The Author(s)</p> EMSA of nuclear proteins from NS and PMA + ionomycin (P/I)-stimulated Jurkat T-cells. () Monoclonal antibodies against Ku were preincubated with nuclear proteins before addition of P-labeled oligonucleotide probe: 162 recognizes the native Ku70/Ku80 heterodimer, 111 recognizes Ku80 and N3H10 recognizes Ku70. Polyclonal antibodies against NF-ATp or cytoplasmic phospholipase A2 (cPLA2) did not inhibit binding of purine-box regulator complex, while cyclosporin A (CsA) caused partial inhibition. () Ku monoclonal antibodies did not inhibit induced NF-κB DNA-binding. () Monoclonal antibodies to NF90 (mAB anti-DRBP76), Ku80 and Ku70, but not NFATc1 or NFATc2, specifically inhibited purine-box regulator ARRE–DNA-binding activity in nuclear extracts of nonstimulated and P/I-stimulated Jurkat T-cells