(A) Daurinol-treated MDA-MB-231 and A549 cancer cells CM was subjected to zymograph assay for analysis of MMP2, MMP9, and uPA activities. Equal volumes of serum-free culture medium were obtained from control cells or cells that had been treated with indicated dose of daurinol for 48 hours. Culture media were concentrated with Amicon Ultra-4 centrifugal device obtained from Millipore and loaded onto gels. The proteolytic activity of MMP-2/MMP-9 and uPA was assessed by SDS-PAGE using zymogram gels containing 0.1% (m/v) gelatin and fibrinogen/plasminogen, respectively. (B) Semi-quantitative RT-PCR was performed for analysis of MMP2, MMP9, and uPA expression. After 48 h of treatment with different concentrations of daurinol in MDA-MB-231 and A549 cancer cells.