FIG. 8.
RelB-p50 fails to induce the Bcl-xL promoter in either SMCs or NIH 3T3 cells. (A and B) Cultures of SMCs at 50 to 70% confluence were transiently transfected, in triplicate, with 0, 0.25, 0.5, 0.75, or 1 μg of RelB expression vector as indicated; 0.5 μg of either the wild-type −298/+22 Bcl-xL-CAT (A) or E8-CAT (B) reporter vector; 0.5 μg of p50 expression 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 two separate experiments. Baseline untreated Bcl-xL and E8-CAT activitywas set at 1, and the relative (Rel.) activities are presented. The results are expressed as mean plus standard error of the mean. (C and D) Cultures of NIH 3T3 cells were transfected as described above with either the wild-type −298/+22 Bcl-xL-CAT (C) or E8-CAT (D) reporter vector, and the relative activities are presented as in panels A and B. (E) Cultures of NIH 3T3 cells were transfected (+) as described above with 0.5 μg of wild-type −298/+22 Bcl-xL-CAT and 0.5 μg of both RelA and p50 expression vectors in the absence or presence of 0.5 μg of CMV IE1 or RelB expression vector plus 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 two separate experiments. Baseline untreated Bcl-xL CAT activity was set at 1, and the relative activities are presented.