Immunohistochemical analysis of aortic tissue

All aortic tissue cross-sections from the above experiments were prepared with a Bond-Max autostainer (Leica Microsystems, Wetzlar, Germany). Slides were stained with primary antibody on a fully automated Bond-Max system and VBS Refine polymer detection system (Leica Microsystems) as previously described (Lo et al., 2017 (link)). Negative controls did not contain primary antibody. Positive immunoreactivity signals in the endothelial layers were measured by ImageJ software. Immunoreactivity signals in endothelial layers from TNF-α and the TNF-α+LSM, TNF-α+LSM+Ab, or TNF-α+rbFGF groups were quantified as previously described (Federici et al., 2002 (link)).