Survivin is transcriptionally regulated by Sp1 in WM cells. (A) BCWM.1 cells were treated with 10 µM of TMP for 24 hours, and purified DNA from chromatin immunoprecipitated with anti-Sp1 and anti-IgG polyclonal antibody was subjected to PCR using primers specific for selected gene promoter regions. The y-axis represents average enrichment over IgG from at least 2 independent experiments normalized to input. (B) BCWM.1 cells with or without TMP treatment at stated concentrations for 24 hours. Cells were then subjected to WB analysis using anti-survivin and anti-GAPDH abs. (C) Survivin expression in BCWM1 and MWCL1 cells transfected with either Sp1-specific siRNA or Scr were analyzed by qRT-PCR. Data were first normalized to GAPDH and then to the signal from control cells. Data are presented as mean of fold change compared with control ± SEM of 3 biological replicates. (D) A plasmid encoding human Sp1, pCAGGS, was transfected in BCWM1 cells using nucleofection. BCWM.1 cells overexpressing Sp1 were evaluated for the expression of survivin by qRT-PCR 2 days after transfection.