A standard procedure was carried out as described previously [36 (link)]. Briefly, coronal sections (8 μm) were prepared at the level of the dorsal hippocampus (1.90–2.50 mm posterior to the bregma). Paraffin sections were dried, washed, permeabilized, blocked in 5% goat serum, and incubated overnight at 4 °C with antibodies against goat anti-SerpinA3N (RD; AF4709), mouse anti-NeuN (Millipore; MAB377), rabbit anti-Olig2 (Abcam; ab109186), rabbit anti-GFAP (Abcam; ab4648), rabbit anti-IBA1 (Huabio; ET1705-78), rabbit anti-phospho-RYR2 (Ser2808) (Biorbyt; orb1093816), and mouse anti-RYR2 (Thermo Fisher Scientific, MA3-916), followed by an Alexa Fluor 680-, 594-, or 488-conjugated secondary antibody (Thermo Fisher Scientific, A-21057, A-11058, A-11001, A-11008) for 1 h at room temperature. Finally, the cells were incubated with mounting medium (with DAPI) (Cat#S2110, Solarbio, Beijing, China) at room temperature for 15 min. Histological images were captured using a Pannoramic MIDI scanner (3D Histech, Hungary). The experiment was repeated in triplicate. ImageJ software was used to manually count positive cells in the same size field of view.
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