Induction of transcriptional suppression of the human
survivin gene by AG490. (A) The survivin promoter, ligated into the SEAP reporter vector pSEAP2–basic to create the construct SEAP2–psurvivin, was transiently transfected into K562 and BC-1 cells. The pSEAP2–basic and the pSEAP2–promoter that contains the simian virus 40 (SV40) early promoter upstream of the SEAP gene were transfected in the same manner. At 24 hours after transfection, cells were suspended at 5 × 105/mL and cultured for additional 48 hours in the presence of AG490 or 0.1% DMSO. The SEAP activities were determined by ELISA. Data represent the mean of triplicate assays. Shown is a representative of 2 experiments performed. TB indicates transcription blocker. (B) The reporter construct psurvivin–EGFP, which contains the survivin promoter upstream of the EGFP gene, was tranfected into BC-1 cells. Cells were treated with 100 μM AG490 or 0.1% DMSO for 48 hours. Live cells were gated by forward (FSC) and side scattering (SSC), and fluorescence intensity was determined on the basis of 5 × 104 live cells analyzed. Shown is a representative of 4 experiments performed.
