Dibutylphthalate polystyrene xylene mounting solution

To block endogenous peroxidase, skin tissue slides were incubated in 0.3% hydrogen peroxide at room temperature for 30 min. The slides were rinsed 3 times with PBS and then incubated in normal animal serum with antibodies overnight to prevent antibody non-specific binding. The slides were then incubated with a proliferating cell nuclear antigen (PCNA) antibody and an ionized calcium-binding adaptor molecule 1 (Iba1) antibody for 12 h and washed three times with PBS. Tissue slides were treated with a biotinylated secondary antibody using the ABC kit (Vector Laboratories Inc, Burlingame, CA, USA) for 2 h, then washed 3 times with PBS. The tissue slides were incubated with 3,3’-diaminobenzidine (Sigma–Aldrich, St. Louis, MO, USA) for 15 min and washed with running water. To stain nuclei, tissue slides were immersed in hematoxylin solution for 1 min, then mounted with a Dibutylphthalate polystyrene xylene (DPX) mounting solution (Sigma–Aldrich). The stained tissues were photographed under an optical microscope (Olympus Optical Co., Korea) and analyzed with the Image J (NIH, Bethesda, MD, USA) software.

To assess status epilepticus-induced neurodegeneration, Fluoro-Jade B (FjB) staining was carried out as before^{51 (link)}. Briefly, 12 μm coronal sections at the medial level of the hippocampus (Bregma AP = − 1.94 mm) were cut on a cryostat. Tissue was fixed in 4% PFA, rehydrated in ethanol, and then transferred to a 0.006% potassium permanganate solution followed by incubation with 0.001% FjB (Chemicon Europe Ltd, Chandlers Ford, UK). Sections were mounted in Dibutylphthalate Polystyrene Xylene (DPX) mounting solution (Sigma Aldrich, Dublin, Ireland). Using an epifluorescence microscope, cells including all hippocampal subfields (dentate gyrus (DG), cornu amonis regions 1 and 3 (CA1 and CA3)) were counted under a 40 × lens in two adjacent sections and the average determined for each animal.