Figure 3: Flunarizine reduces Angiopoietin-2 (Angpt-2) synthesis in vitro.
From: Flunarizine suppresses endothelial Angiopoietin-2 in a calcium - dependent fashion in sepsis
(A) Cropped Angpt-2 immunoblot from human umbilical vein endothelial cell (HUVEC) lysates 15 hrs after 10 μM Flunarizine (FLU) (+) or vehicle (−) treatment. (n = 4) (B) After stimulation with 10 μM FLU or vehicle for 1 h, 10 ng/mL Tumor necrosis factor α (TNFα) or control was applied to HUVECs for 24 hrs. Angpt-2 concentration in cell lysates was measured by enzyme-linked immunosorbent assay (ELISA) and is shown relative to whole protein (n = 6). (C) Fluorescent immunocytochemistry for Angpt-2 (red) and nuclear staining (4′,6-diamidino-2-phenylindole, blue) in HUVECs after treatment with 10 μM Flunarizine, vehicle or Angpt-2 siRNA as a negative control for 24 hrs (n = 4) (D) Real-time polymerase chain reaction (RT-qPCR) for Angpt-2 in HUVECs stimulated with 10 ng/mL TNFα or control for 15 hrs after pretreatment with either FLU or vehicle (n = 6) for 1 h. Columns are presented as mean ± SEM.
