(A) Cells were treated with 100 μM AG490 or 0.1% DMSO for 24 hours, and equal amount of cell lysates (25 μg per lane) were subjected to SDS-PAGE. Western blotting was performed to detect Bcl-2, Bcl-X_L, Mcl-1, and survivin protein. Data shown are representative of 3 independent experiments. (B) Kinetic analysis of survivin protein expression. Cells were treated with 100 μM AG490 for the indicated time periods and analyzed for expression of survivin protein. Data shown are representative of 2 independent experiments. (C) Dominant-negative STAT3 reduces survivin expression. Cells were transfected with plasmid vectors for expression of EGFP-STAT3D or EGFP. At 2 days later, EGFP⁺ cells were sorted and lysed by RIPA buffer, and equal amounts of protein (30 μg per lane) were subjected to SDS-PAGE. Western blotting was performed for STAT3, survivin, and actin. Data shown are representative of 2 independent experiments. Ctr indicates control; DN, dominant-negative STAT3.