FIG. 1.
IE1 fails to induce Bcl-xL promoter activity in SMCs and NIH 3T3 cells. (A) Cultures of SMCs at 50 to 70% confluence were transiently transfected, in triplicate, with 0, 0.25, 0.5, or 1 μg of pON2205 vector expressing IE1 protein, 0.5 μg of either the wild-type −298/+22 Bcl-xL-CAT (open bars) or −298/+22 mkB Bcl-xL-CAT (solid bars) Bcl-xL reporter vector, 0.5 μg of SV40 β-Gal expression vector, and enough pBluescript to make a total of 2.5 μg of DNA. After 48 h, the cells were harvested and CAT activity and β-Gal activity were measured. The data presented are the mean of four separate experiments. Baseline untreated mutant Bcl-xL promoter CAT activity was set at 1. The results are expressed as mean plus standard error of the mean. (B) Cultures of NIH 3T3 cells were transfected and processed as described for panel A. (C) Cultures of NIH 3T3 cells were transfected, in triplicate, with 0, 0.25, 0.5, 0.75, or 1 μg of RelA expression vector, 0.5 μg of p50 expression vector, 0.5 μg of wild-type −298/+22 Bcl-xL-CAT, 0.5 μg of β-Gal expression vector, and enough pBluescript to make a total of 2.5 μg of DNA and processed as described for panel A. Baseline untreated wild-type Bcl-xL promoter CAT activity was set at 1.