Fig. 5.

Lentivirus-mediated reexpression of TM in TM^−/− cells, but not sTM, normalizes VWF and Ang2 levels. (A) Levels of VWF, Ang2, and TM RNA transcripts in WT, TM^−/−, and TM^match were measured by qRT-PCR. GAPDH was used as an internal control for quantitation. (B) Confluent WT, TM^−/−, and TM^match cells were lysed, and levels of VWF, Ang2, and β-actin were analyzed by Western blotting. Quantitation of changes in the levels of (C) VWF and (D) Ang2 in WT, TM^−/−, and TM^match cells. Levels of (E) VWF and (F) Ang2 in the medium supernatant collected from confluent WT, TM^−/−, and TM^match cells. (G) Levels of VWF in the medium supernatant collected from confluent WT and TM^high cells were measured by a sandwich ELISA. (H) Confluent WT and TM^high cells were lysed, and levels of VWF, TM, and β-actin were analyzed by Western blotting. (I) Confluent WT and TM^−/− cells were treated with sTM (100 nM) for 16 h in the basal medium containing 0.5% BSA, and levels of VWF in the medium supernatant were analyzed by a sandwich ELISA. Data are mean ± SEM (n = 3). One-way ANOVA: *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.001.