FIG 2.

U54 inhibits NFAT transcriptional activity. (A) 293T cells were transfected with 4TO, 4TO-U54, 4TO-U11 and NFAT-Luc reporter plasmids. Cells were stimulated for 24 h or left unstimulated, and luciferase activity was determined and normalized to protein content (n = 9). Western blot analysis confirmed the expression of proteins of interest for each condition tested. U54-Myc and U11-FLAG proteins were expressed in cells that had been transfected with 4TO-U54 and 4TO-U11 plasmids (top). Beta-actin was included as a loading control. (B to D) Transcriptional activities of AP-1 (B), CRE (C), and NF-κB (D) were evaluated in 293T cells by luciferase assay. 293T cells were transfected with 4TO, 4TO-U54, pZVH14, IE2, and either pAP1-Luc, pCRE-Luc, or pNF-κB-Luc. Luciferase activity was determined by luciferase assay and normalized to a BCA assay (n = 3).