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. 2014 Nov;88(21):12452–12463. doi: 10.1128/JVI.02030-14

FIG 4.

FIG 4

U54 inhibits NFAT1 and NFAT2 activity on the IL-2 promoter. Transcriptional activity of ectopically expressed NFAT1 and NFAT2 factors was evaluated in 293T cells by luciferase assay. (A) 293T cells were transfected with 4TO, 4TO-U54, 4TO-U11, REP-NFAT1, REP-NFAT2, and IL-2-Luc reporter plasmids. Cells were stimulated for 24 h or left unstimulated, and then 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 or U11-FLAG proteins were expressed in cells transfected with 4TO-U54 or 4TO-U11 (upper panels), and NFAT1 or NFAT2 proteins were expressed in those transfected with REP-NFAT1 or REP-NFAT2 (middle panels). Beta-actin was included as a loading control. (B) 293T cells were transfected with 4TO, 4TO-U54, 4TO-U54A, REP-NFAT1, and IL-2-Luc reporter plasmids. Cells were stimulated for 24 h or left unstimulated, and luciferase activity was determined by luciferase assay and normalized to protein content (n = 6).