Figure 2. Loss of BMPRII induces endothelial inflammation by mechanisms dependent on ROS, NADPH oxidase, and NFκB activity.
(A, B) HUVECs transfected with BMPRII siRNA or Non.si were also treated with apocynin (60 μM, 2 days) or vehicle, and monocyte adhesion assay (A) or Western blot analysis (B) of cell lysates using ICAM-1 antibody were performed (mean ± SEM, n=4, *p<0.05). (C, D) Thoracic aorta sections of BMPRII+/+ApoE−/− and BMPRII+/− ApoE−/− mice were stained with dihydroethidium for ROS detection (C) or Nox1 antibody (D). Shown are representative confocal microscopy images (n=6 each). The insets show magnified views of endothelial regions. Nuclei (blue) and elastic laminas (green) are shown. L: lumen. Also shown are en face confocal images of Nox1 antibody staining (lower panels, D). (E) HUVECs were transfected with Non.si, BMPRII siRNA, BMPRII siRNA+Nox1 siRNA, or Nox1 siRNA for 2 days and analyzed by Western blots using ICAM-1, VCAM-1, BMPRII, Nox1 and β-actin antibodies. (F-H) HUVECs transfected with BMPRII siRNA or Non.si were treated with Bay11-7082 (10μM, 24 h) before monocyte adhesion assay (F), qPCR analysis for BMPRII, ICAM-1 and VCAM-1 (G), and Western blots with BMPRII, ICAM-1 and VCAM-1 antibodies (H), (n=4, *p<0.05).