Immunofluorescent Staining of Aortic Tissue

The detail of immunofluorescent staining was described in our previous study [17 (link)]. Briefly, the OCT-embedded aorta root was cut into sections at 6 μm using a Lab-Tek tissue processor Leica CM1950. Sections were incubated with anti-Mac-2 (1 : 100, #CL8942AP, Cedarlane) and anti-p65 (1 : 200 dilution, Thermo Fisher Scientific) at 4°C overnight, followed by being stained with Alexa Flour 555 (1 : 300, #A21434, Invitrogen) or 488 (1 : 300, #A11034, Invitrogen) for 1 hour at room temperature. DAPI (#P36935, Invitrogen) was used to label the nuclei. All images were collected by confocal laser scanning microscopy (Nikon, Japan).