Integration of flow-dependent endothelial phenotypes by Kruppel-like factor 2
J. Clin. Invest. Kush M. Parmar, et al. 116:49 doi:10.1172/JCI24787 [Go to this article.]

Figure 3
KLF2 regulates genetic programs controlling blood vessel development, vascular tone, and thrombosis/hemostasis. (A) HUVECs under the same conditions as in Figure 2A were analyzed by quantitative TaqMan RT-PCR (n = 3) of the indicated genes; Ad-GFP was performed as the control. (B) Ang-2 protein levels in supernatants were measured by ELISA under the same conditions (n = 3) as in A, but with an 8-hour incubation with IL-1β. Ang-2 levels shown are after subtraction of the amount in the media before conditioning. (C) Western blot of Tie2 and GAPDH (loading control) on whole-cell lysate of HUVECs infected with Ad-GFP or Ad-KLF2 for 24 hours. (D) Quantitative TaqMan RT-PCR was performed to assess the levels of expression of eNOS, ASS, CNP, and ET-1. (E) CNP protein levels in supernatant measured by ELISA. Supernatant was collected from HUVECs infected with the indicated adenoviruses for 24 hours with or without IL-1β. nd, not detectable. (F) Effect of KLF2 and/or IL-1β on the expression of major genes involved in thrombosis. Quantitative TaqMan RT-PCR of indicated genes was performed as described in A. EDG-1, endothelial differentiation gene 1; PAI-1, plasminogen activatory inhibitor 1. (G) FACS analysis of surface TM (TM/CD141) expression on HUVECs infected with Ad-GFP or Ad-KLF2 for 24 hours. (H) FACS analysis of cell-surface TF expression on HUVECs infected with the indicated virus followed by incubation with media or IL-1β (10 U/ml) for 6 hours. Bar graphs represent mean ± SEM (n = 3). *P < 0.05, **P < 0.01 vs. control; Student’s t test.