Tissue sections at 4μm in each animal group were used to perform immunostaining. Incubated with the primary antibody at 4°C and then with a horseradish peroxidase-conjugated secondary antibody. Color was developed by incubating with diaminobenzidine (DAB Detection Kit, GK600505; Gnen Tech, Shanghai, China). CD31 and BrdU double staining was performed using VECTASTAIN ABC-AP Kit, alkaline phosphatase (Mouse IgG) (AK-5002, Vector Labs, Peterborough, UK) and Vector®Red (SK-5100, Vector Labs) following the manufacturer’s instructions. Hematoxylin was then used to stain the nucleus. The photos were scanned with a digital slide scanner (Pannoramic SCAN, 3DHISTECH).

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