1. Sp1 complementation assay.
Sp1 complementation assay. A. U2OS cells were transfected with nontargeting control or Sp1 siRNA. Seventy-two hours after siRNA transfection, cells were transfected with Sp1WT or Sp1S101A. Forty-eight hours after transfection, cells were exposed to the indicated amount of H2O2 for 1 h. Cells were then plated at a density of 300 cells per 60-mm dish. After 11 d, discrete colonies were stained with crystal violet and counted in a blinded manner. Data are expressed relative to the number of colonies in the nontreated cells receiving the same DNAs. B. U2OS cells were exposed to the indicated concentrations of H2O2 for 1 h and harvested directly in SDS sample buffer. Immunoblot was done with antibody specific to HA.11 and γSp1101. Bottom inset, Western blot shows Sp1 (top) and nucleolin (bottom) levels 72 h after transfection with Sp1 or nontargeting control siRNA.
Beatrix A. Olofsson et al. Mol Cancer Res 2007;5:
©2007 by American Association for Cancer Research