PDI silencing affects procoagulant activities on ECs. (A) Analysis of basal and ionomycin-decrypted TF procoagulant activity in response to transient PDI silencing and/or exogenous hPDI addition. TF activity is normalized to TF antigen and represented as fold increase over nontransfected ECs. (B) Analysis of basal and ionomycin-enhanced prothrombinase activity in response to transient PDI silencing and/or exogenous hPDI addition. The prothrombinase activity is normalized to the total protein content and represented as fold increase over nontransfected ECs. (C) Representative Western blot and densitometric analysis of PDI in puromycin-selected PDI shRNA-tranfected TF-EC lines. (D) Comparative analysis of basal TF procoagulant activity (open bars) and prothrombinase activity (solid bars) of PDI shRNA-transfected TF-EC cell lines shown in panel C. Procoagulant activities were normalized to nontransfected TF-ECs. (A-B,D) Data are mean ± SD of triplicates. Statistical analysis was performed by 1-way ANOVA: *P < .05, **P < .01, ***P < .001.