Flow Cytometry Analysis of Aortic Cells

For flow cytometry analyses, single-cell suspensions of mouse aortae were prepared according to the previous protocol (20 (link)) and stained with following antibodies: 7AAD (00-6993-50, eBioscience, USA), Fluorescent isothiocyanate (FITC) anti-mouse-α-SMA (ab8211, Abcam, United Kingdom), PE-anti-mouse-CD34 (551387, BD, USA), FITC-anti-mouse-CD68 (MA1-82739, Invitrogen, USA), PE-anti-mouse-NK1.1 (ab269324, Abcam, United Kingdom), FITC-anti-mouse-CD19 (11-0193-82, eBioscience, USA), PE-anti-mouse-CD21 (12-0211-82, eBioscience, USA), PE-anti-mouse-CD45 (12-0451-82, eBioscience, USA), and Collagen I (ab88147, Abcam, United Kingdom; at room temperature for 30 min) with Alexa Fluor 488 goat anti-mouse antibody (A32723, Invitrogen, at room temperature for 30 min). Flow cytometry results were analyzed by using FlowJo 7.6 software (FlowJo, LLC., Ashland, Oregon). Statistical analysis was further performed by GraphPad Prism 8.0.