Figure 1. Identification and expression of tissue factor pathway inhibitor after gene transfection. A: Restriciton enzyme analysis of TFPI-2 plasmids digested with XbaI and KpnI. Line 1 is the 2-Log Marker. Line 2 is the pBos-Cite-neo/TFPI-2. Line 3 is the pBos-Cite-neo/TFPI-2 digested with XbaI and KpnI. Line 4 is the λDNA/EcoRI+HindIII Markers. B: Semi-quantification of TFPI-2 mRNA expression in hTCFs after transfection with an empty vector or TFPI-2 expression vector was assessed by RT-PCR. RT-PCR for β-actin mRNA was performed as a control. Relative OD Value (OD TFPI-2/OD β-actin) was used to evaluate the expression level of TFPI-2. Data are presented as mean value±SD, the double asterisk indicates a p<0.001, Student’s t-test. C: Western blot analysis for the expression of TFPI-2 in hTCFs after transfection with an empty vector or TFPI-2 expression vector. Each protein fraction was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis on 15% acrylamide gels, transferred onto nitrocellulose membranes, and probed with a rabbit anti-human TFPI-2 antibody. Molecular sizes of the detected bands are shown to be 33, 31, and 27 kDa.