Effect of NF-κB, CREB, CBP, and p300 on TIPS. Jurkat-LT cells were plated in 12-well tissue culture dishes at 1 × 106cells/well and 1.5 μg plasmids was transiently transfected using SuperFect transfection Reagent (Qiagen). Wild-type SHP-1 P2 promoter (wt-P2-Luc, 500 ng) was cotransfected with increasing amounts of p65 (RelA) or p50 of NF-κB family (0, 30, 100, and 300 ng), CREB (0, 25, 50, 100 ng), CBP (0, 10, 50, 100 ng), or p300 (0, 10, 50, 100 ng) in the absence (A) or presence of 200 ng wt Tax (B), M22 Tax (C), or M47 Tax (D). (E) Wild-type SHP-1 P2 promoter (wt-P2-Luc, 500 ng) was cotransfected without or with 30 ng RelA plasmid and 100, 200, or 400 ng wt, M22, or M47 Tax. Cell lysates were assayed in triplicates for luciferase activity and values represent the mean ± SD of 2 experiments. The relative luciferase activity was normalized against basal SHP-1 P2 promoter luciferase reporter alone (set as 100).
