Gb13428

A fluorescent double staining kit (G1235; Servicebio, Wuhan, China) based on tyramide signal amplification was performed in accordance with the manufacturer’s instructions. Primary antibodies for histological staining included CD31 (dilution 1 : 5000; GB13428; Servicebio) and PDGFR‐β (dilution 1 : 500; #3169; Cell Signaling, Danvers, MA, USA).
Purified pericytes were fixed, permeabilized, and subsequently blocked with BSA for 1 h. Cells were then incubated with antibodies against α‐SMA (dilution 1 : 100; ab7817; Abcam, Cambridge, UK), PDGFR‐β (dilution 1 : 100; #3169; Cell Signaling), neuron‐glial antigen 2 (NG2) (dilution 1 : 100; sc‐53389; Santa Cruz Biotechnology, Dallas, TX, USA), and CD13 (dilution 1 : 100; sc‐13536; Santa Cruz Biotechnology) overnight at 4 °C. The next day, the cells were washed and incubated with secondary Alexa Fluor 488 anti‐rabbit antibody (dilution 1 : 1000; 4412; Cell Signaling) or Alexa Fluor 594 anti‐mouse antibody (dilution 1 : 1000; 8890; Cell Signaling) for 1 h when protected from light. Nuclear staining was performed with 4′,6‐diamidino‐2‐phenylindole. The images were captured by high‐content analysis (Operetta; PerkinElmer, Waltham, MA, USA). Image analysis and fluorescence quantification were performed using columbus (PerkinElmer) and imagej (NIH, Bethesda, USA).

Sections were incubated with anti-CD34 (AF5149, Affinity Biosciences, USA), anti-CD133 (AF5120, Affinity Biosciences, USA), and anti-CD31 (GB13428, ServiceBio, China) at 4°C overnight. Then, sections were immunoblotted with fluorescence-conjugated secondary goat anti-rabbit antibodies. The nucleus was stained with 4’6-diamidino-2-phenylindole (DAPI). Finally, images were captured under a fluorescence microscope.