Fig. 8

In vivo deletion of endothelial Notch1 results in inflammatory cell recruitment. a Timeline for inducible deletion of endothelial Notch1 to indicate animal ages and evaluation time points. b Quantification of CD68 expression from the intima of descending aortae of adult WT mice (n = 9) and Notch1 ^ECKO mice (n = 5) 2 weeks post tamoxifen by qPCR analysis. Graph bars represent average ± SEM, T-test *P < 0.01. c Confocal tile scan of the descending aorta of Notch1 ^ECKO animal at 8 weeks post tamoxifen reveals the presence of CD45⁺ tdTomato⁻ cells embedded within the tdTomato⁺ endothelium. Scale bar = 500 µm. d Ve-Cadherin and CD45 staining of control (Cre⁻ Notch1 ^loxP/loxP) and Notch1 ^ECKO descending aortae at 8 weeks post tamoxifen displays CD45⁺ inflammatory cells at higher magnification in the Notch1 ^ECKO aorta; scale bar = 10 µm. e En face imaging of the endothelium by Ve-Cadherin staining in the descending aorta of control (Cre⁻ Notch1 ^loxP/loxP) and Notch1 ^ECKO mice, both tamoxifen-injected and sacrificed after 8 weeks, shows proliferating (EdU⁺) CD45⁺ inflammatory cells (marked by open arrow heads) embedded within the endothelial layer of Notch1 ^ECKO mice; scale bars = 10 µm. f Three-dimensional surface render to visualize the spatial location of marked CD45⁺ cell relative to the endothelium (marked EC) and smooth muscle cells (marked SMC). Both bottom and side view reveal that the CD45⁺ cell is locating above the smooth muscle cell layer. Scale bars = 10 µm