Research Paper Volume 5, Issue 7 pp 515—530

Figure 1. Endothelium-specific overexpression of Nox2 and ROS generation. (A) Schematic used to make conditional binary transgenic mice (NVF). Tet-ON, tetracycline in the drinking water to suppress the transgene; Tet-OFF, withdrawal of tetracycline for four weeks to induce the transgene. (B) Frozen heart sections of 8 weeks old Tet-ON and Tet.OFF NVF (for 4 weeks) mice showing EC-specific (CD31, green) HA-tagged Nox2 (red) expression in coronary vessels of Tet-OFF animal. (C) Immunohistochemistry using anti-HA antibody on frozen heart sections. (D) Western blots using transgenic mouse heart EC (MHEC) lysates showing Nox2 overexpression in two independent lines of Tet-OFF animals compared to Tet-On and WT animals. (E) Q-PCR using MHEC RNAs from Tet-ON and Tet-OFF animals (n=6/group). (F) FACS analyses using DCF fluorescence of MHECs (n=6/group). All animals were 8 weeks old, Tet-OFF were without tetracycline for four weeks. Tet-ON MHECs were grown in medium containing tetracycline 2μg/mL. *p<0.05.