FIG 3.

U54 inhibits ectopically expressed NFAT1 and NFAT2 transcriptional activity. (Top) Transcriptional activity of ectopically expressed NFAT1 and NFAT2 factors was evaluated in 293T cells by luciferase assay. 293T cells were transfected with 4TO, 4TO-U54, 4TO-U11, REP-NAFT1, REP-NFAT2, and NFAT-Luc reporter plasmids. Cells were then stimulated for 24 h or left unstimulated, and luciferase activity was determined and normalized to protein content (n = 9). (Bottom) Western blot analysis confirmed the expression of proteins of interest for each condition tested. U54-Myc or U11-FLAG proteins were expressed in cells that had been transfected with 4TO-U54 or 4TO-U11 plasmids (upper panels), and NFAT1 or NFAT2 proteins were expressed in those transfected with REP-NFAT1 or REP-NFAT2 plasmids (middle panels). Beta-actin was included as a loading control.