Sections (4 μm) were cut from the paraffin-embedded aortic specimens obtained from the aortic dissection patients, non-ruptured aneurysm patients, healthy body donation volunteers or mice. Human aortic sections were immunostained with rabbit polyclonal antibody against human glucocorticoid receptor (GCR, 1:100, Abcam, ab3580, Manassas, VA, USA), and mouse monoclonal antibody against human macrophage (CD 64, 1:150, Abcam, ab140779). The negative control slides were stained with control IgG (Supplemental Fig. II). Blind evaluation of positive staining percentages was performed in 10 different fields at 400 × magnification from two independent pathologists, with an intervariability < 5%.
Mouse aortic tissues were stained with hematoxylin and eosin (HE) and Masson's trichrome staining. Photomicrographs were analyzed by investigators blinded to the experimental protocol using image pro-plus software to assess aortic media thickness and collagen volume fraction. Aortic dissection was confirmed by HE photograph, which was defined as the coexistence of true lumen and false lumen, even the thrombus in false lumen. They were also immunostained with rabbit polyclonal antibody against GCR (1:100, Abcam, ab3580), rat monoclonal antibody against macrophage (F4/80, 1:20, Abcam, ab6640) or control IgG.
Mouse aortic tissues were stained with hematoxylin and eosin (HE) and Masson's trichrome staining. Photomicrographs were analyzed by investigators blinded to the experimental protocol using image pro-plus software to assess aortic media thickness and collagen volume fraction. Aortic dissection was confirmed by HE photograph, which was defined as the coexistence of true lumen and false lumen, even the thrombus in false lumen. They were also immunostained with rabbit polyclonal antibody against GCR (1:100, Abcam, ab3580), rat monoclonal antibody against macrophage (F4/80, 1:20, Abcam, ab6640) or control IgG.