Pe anti mouse cd34

Flow cytometry was performed for analysis of cell surface antigen expression. The MSCs at passage one were detached using Accutase (Innovative Technologies Inc., San Diego, CA, USA) and resuspend into PBS at a concentration of 1 × 10⁵ cells per 100 µL. The cells were then incubated with fluorescence-labeled primary antibody or isotype control for 30 min in dark condition. The utilized primary antibodies were: FITC anti-mouse Ly-6A/E Sca-1 (Biolegend), PE anti-mouse CD 90, PE anti-mouse CD44, PE anti-mouse CD 146, FITC anti-mouse CD14, PE anti-mouse CD34 (BD, Franklin Lakes, NJ, USA). These samples were analyzed by Accuri™ C 6 Flow Cytometer (BD).

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.