FIG. 7.

Mapping of promoter elements required for PI3-K/Akt and IL-3 activation of the mcl-1 reporter gene. (A) Ba/F3 cells cotransfected with the indicated reporter gene and an empty expression vector (−Akt*) or vector encoding M-Akt (+Akt*) were left in medium without IL-3 for 12 h before cell lysates were analyzed for luciferase activity. (B) Same as panel A except that cells were transfected with the indicated reporter construct alone and were left untreated (−IL-3) or stimulated with IL-3 (+IL-3) for 12 h. (C) Ba/F3 cells transfected with the reporter plasmids as indicated plus an expression vector encoding nothing (Vector), DNAkt, or DNp85 were starved and restimulated with IL-3 for 3 h before cell lysates were prepared and analyzed for luciferase activity. The results shown are averages from five independent transfection assays and are plotted as relative activities to cells transfected by the same reporter plasmid and deprived of IL-3. The latter activity was considered to be 100. Numbers inside the bar graph are P values (calculated by the Student t test) for various conditions as indicated.