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. 2002 Jan;76(1):232–242. doi: 10.1128/JVI.76.1.232-242.2002

FIG. 3.

FIG. 3.

EBNA-3C coactivation of the LMP1 promoter element is independent of RBP-Jκ. (A) Representative assay (of two repetitions) using BJAB cells transfected with the reporter plasmid p(−236/−145)LMP1p-Luc containing the −236/−145 LMP1 promoter element upstream of a minimal SV40 early promoter and the luciferase ORF, along with the indicated amounts of pSG5-EBNA-2 (E2) and pSG5-EBNA-3C (E3C). Fold increase in luciferase activity is indicated. (B) BJAB cells were transfected with CAT reporter plasmids containing wild-type (wt) or mutant LMP1 promoter sequences and the indicated amounts of pSG5-EBNA-2 and pSG5-EBNA-3C. Results are representative of two independent repetitions. Percent conversion was determined using ImageQuant software and a PhosphorImager. Fold activity is relative to that of the vector-only control, which was assigned a value of 1. (C) BJAB cells were transfected with p(−512/+72)LMP1p-Luc and the indicated amounts of pSG5-EBNA-2-SS (E2-SS) and pSG5-EBNA-3C. Results are representative of independent duplicate experiments. Fold activation of luciferase is indicated. (D) BJAB cells were transfected with p(−512/+72)LMP1p-Luc and the indicated amounts of pSG5-EBNA-2 and pSG5-flagEBNA-3C, pSG5-flagEBNA-3A, or pSG5-flagEBNA-3B. The results are representative of two independent experiments. Relative luciferase activity is indicated. The panel below the graph shows a corresponding immunoblot with monoclonal antibodies directed against the Flag epitope tag. WB, Western blot.