Figure 5.

Micro-RNA (miR)-126 reduces the expression of both alternatively spliced tissue factor (asTF) and full-length (fl) TF in human microvascular endothelial cells (HMEC) after stimulation with tumor necrosis factor (TNF)-α. HMECs were cultured for 24 h and then transfected with a control mimic or a miR-126 mimic (A–C) as well as an inhibitor control or anti–miR-126 (D–F). The transfection efficiency was confirmed by real-time TaqMan polymerase chain reaction (A and D). Twenty-four hours post transfection, the cells were stimulated with 10 ng/mL TNFα. Gene expression was determined for asTF (B and E) and flTF (C and F) 2 h after stimulation with TNFα. GAPDH was used as house-keeping gene. Data are represented as mean±SEM. *P<0.05, **P<0.01, ***P<0.0001, n≥3. Representative western blots show the protein expression of flTF and asTF after transfection with miR-126 (G) or anti–miR-126 (H) 6 h post TNFα stimulation. GAPDH was used as loading control (n=3).