5.

Blocking EPCR prevents APC-induced MMP-2 synthesis and proliferation. (A) ECPR protein expression in response to EPCR siRNA treatment at 48 hrs detected by Western blot using whole cell lysates. β-actin was used as the loading control. (B) Cell proliferation was measured after cells were treated with combinations of APC, siRNA and RCR252 for 72 hrs. Cell proliferation is expressed as a percentage of control (mean ± S.D., n= 3 experiments). *P < 0.05 and **P < 0.01 compared to negative control (first bar), ⁺⁺P < 0.01 compared to APC (fourth bar). Images of gels represent one of three independent experiments. (C) MMP-2 was measured in supernatants of cells trans-fected with either control siRNA (0) or EPCR siRNA for 72 hrs, using zymography. (D) MMP-2 was measured in supernatants of cells treated with APC alone or APC plus RCR252 for 24 hrs using Western blot analysis. β-actin used as the loading control.