Brain sections were blocked for 1 hour at room temperature with 5% BSA and incubated with the primary antibodies including mouse anti-GFAP (1:1000) and rabbit anti-AQP4 (1:400) at 4 °C overnight. The next day, all sections were rinsed 3 times and incubated for 2 hours at room temperature in a mixture of Alexa Flour™ 555 donkey anti-mouse IgG (1:1000; Thermo Fisher; Cat. A31570) and Alexa Flour™ 488 donkey anti-rabbit IgG (1:1000; Thermo Fisher; Cat. A21206). After rinsing, the sections were incubated for 6 min in 4',6-diamidino-2-phenylindole (DAPI) (1:1000; Thermo Fisher; Cat. D21490) and coverslipped with antiuorescent quencher.
Thio avine-S staining Depara nized sections were incubated with 1% Thio avine-S (Sigma-Aldrich) for 5 min. Next, 70% ethanol was used to differentiate for 5 min, followed by rinsing with distilled water. The brain sections coverslipped with anti-uorescent quencher.
Thio avine-S staining Depara nized sections were incubated with 1% Thio avine-S (Sigma-Aldrich) for 5 min. Next, 70% ethanol was used to differentiate for 5 min, followed by rinsing with distilled water. The brain sections coverslipped with anti-uorescent quencher.