Af5164

The reagents used in the experiment were aloe-emodin (BJS9141396, Picasso, Shanghai, China), fluoxetine hydrochloride (J20130010, Patheon, Suzhou, China), SDS-PAGE gel preparation kit (G2003-50T, Servicebio, Suzhou, China), PVDF membrane (IPVH00010, Millipore of USA MA), Allergic ECL Chemiluminescence substrate (BL520B, Biosharp, Anhui, China), Beta Actin-Ab (AF7018, Affinity, Jangsu, China), goat anti-rabbit secondary antibody -HRP (70-GAR0072, Multi sciences, Hangzhou, China), GAPDH antibodies (AF7021, Affinity, Jangsu, China), AQP3 antibodies (AF5222, Affinity, Jangsu, China), AQP4 antibodies (AF5164, Affinity, Jangsu, China), AQP5 antibodies (AF5169, Affinity, Jangsu, China), BDNF antibodies (ab108319, Abcam, Shanghai, China).

After embedding in paraffin wax, the tissues were sliced to 4 μm thickness, then incubated in 0.3% H₂O₂ for 30 minutes and incubated in 0.1% Triton X-100 for 20 minutes. Next, the sections were incubated with primary antibodies, including anti-Aquaporin-4 (AQP4) antibody (1 : 50; AF5164; Affinity) and anti-claudin-5 antibody (1 : 50; AF5216; Affinity) overnight at 4°C and secondary antibody for 60 min at 37°C. Lastly, the sections were stained with DAB to develop the color and counterstained with hematoxylin.