Briefly, after being anesthetized with isoflurane and intracardially perfused with PBS and 10% formalin, brains were removed, fixed in 10% formalin overnight, immersed in 30% sucrose for 3 d, and then frozen. Fixed frozen brain tissues were sectioned at 10 μm by using cryostat (Leica CM3050S-3-1-1, IL). Double immunofluorescence staining was performed as previously reported (He et al. 2015 (link)). Sections were permeabilized with 0.3% Triton X-100 for 30 min. After blocking with 5% donkey serum for 1h, the sections were incubated with the following primary antibodies: anti-MSP (1:1000, Abcam, MA, RRID: AB_10976147), anti-RON (1:1000, Abcam, MA, RRID: AB_10972503), anti-GFAP (1:1000, Santa Cruz Biotechnology, TX, RRID: AB_627673), anti-vWF (1:500, Santa Cruz Biotechnology, TX, RRID: AB_10842026) and anti-NeuN (1:500, Abcam, MA, RRID: AB_2532109), at 4℃ overnight, followed by appropriate fluorescence-conjugated secondary antibodies (Jackson ImmunoResearch Laboratories, PA, RRID: AB_2337972, AB_ 2338059, AB_2340432 or AB_2338871) at room temperature for 2h. The co-localizations were evaluated by a fluorescent microscope (Olympus OX51, Tokyo, Japan).