A32766 a 21203 a 21206 a32754

Tissue sections were permeabilized with 0.5% Triton X-100 in PBS solution and closed with 5% BSA for 1 h. Cells were then fixed with 4% paraformaldehyde at room temperature for 15 min, followed by two washes in PBS and 0.5% Triton X-100 (P0096, Beyotime, China) permeabilized for 10 min. Tissue sections or cells were then mixed with primary antibodies: Rabbit anti-MAP2 (ab183830, 1:500), rabbit anti-GFAP (ab207165, 1:500), rabbit anti-SYN1 (#5297, 1:200, CST, USA), mouse anti-Nestin (ab81462, 1:100), rabbit anti-SOX2 (ab92494, 1:100), rabbit anti-F4/F80 (ab6640, 1:200) were incubated overnight at 4°C. After washing three times with PBS solution. Next, the sections were incubated with Alexa Fluor 488/594-coupled secondary antibodies (A32766/A-21203/A-21206/A32754, 1:500, Thermo Fisher Scientific, USA) for 1 h. Then, the sections were washed three more times with PBS solution and incubated with DAPI The sections were stored at 4°C and then observed with a confocal microscope (Leica SP8).^{32 (link)}
Primary antibodies were purchased from Abcam (UK), except for SYN1.