A region of 1100-nt upstream of pri-miR-146a is able to induce expression of a reporter gene upon TCR stimulation. Jurkat cells were cotransfected with pMluc luciferase reporter containing the 1100-bp miR-146a promoter region and with pGL3-tk-luc, used for normalization of transfection. A renilla/firefly Dual-Luciferase assay was performed. Relative promoter activity in stimulated cells is compared with that in nonstimulated cells, which is set to 1. (A) Cells were stimulated for 24 hours with either PMA and ionomycin or anti-CD3/CD28 Abs. Where indicated, Jurkat cells were pretreated for 30 minutes with CsA. Each bar is the mean of 4 independent experiments (P < .05). (B) Jurkat cells were cotransfected with the wild-type or with the mutagenized 1100-bp promoter constructs: NFKB(1); NFKB(2); c-ETS; or USF-2 together with pGL3-tk-luc, used for normalization of transfection. Cells were stimulated with anti-CD3/CD28 Abs for 24 hours. Each bar is the mean of 4 independent experiments (P < .05).