CD4+ T cells gene-transduced with AML1, wild type Foxp3, and a Foxp3 mutant defective in binding to AML1

To clarify how Foxp3 regulates its target genes, we performed co-immunoprecipitation experiments and found that Foxp3 physically bound to AML1/Runx1 (Ono, M. et al, Nature, 2007). In this series of study, we compared gene regulations by AML1, wild type Foxp3, and a Foxp3 mutant with defective binding to AML1. Keywords: Retroviral gene-transduction into primary CD4+ T cells CD4+ naive T cells were activated and retrovirally gene-transduced with either empty vector (pMCsIg), or AML1-, wild type Foxp3-, or an AML1-non binding mutant Foxp3-encoding vectors. Sixty hours after transfection, 5 x 10^6 GFP-expressing cells were sorted and total RNA was extracted. One cycle target labelling protocol was used for labelling RNA with biotin.