After 4% PFA post-fixation, brains were cut with a vibratome (Leica microsystem) in serial free-floating coronal sections of 35-µm thickness.
The brains were cut with a cryostat in the coronal plane from the level of the tractus septopallio-mesencephalicus to the caudal end of the tuberal hypothalamus.
Brains were cut with a vibratome in coronal corticostriatal slices (250 µm) in ice-cold solution, whose composition was (in mM): 110 choline, 2.5 KCl, 1.25 NaH2PO4, 7 MgCl2, 0.5 CaCl25 25 NaHCO3, 7 glucose, pH = 7.4 bubbled with O2/CO2 (95/5%).
For comparing gene expression patterns of the PRMS and DRMS, freshly dissected brains were cut with a vibratome (Leica, Wetzlar, Germany) in 100 μm sections, and the corresponding brain regions were isolated using a binocular microscope and shock frozen immediately after isolation.
The brains were cut with a coronal section at the injection site, and the anterior part was mechanically disrupted and used for establishing cell cultures whereas the posterior part was formalin-fixed and used for paraffin sectioning.
Coronal sections of the brain were cut at a 35 μm thickness with a cryosectioning microtome and stored at 4 °C in PBS containing 0.1%% sodium azide until use.
Two brain tissue samples around the hematoma of each brain were cut and weighed.
30-μm thick brain sections were cut with a cryostat (CM3050, Leica) and kept floating in PB 0.1 M pH 7.4.
The brain was removed from the skull and post-fixed overnight in 4% PAF-PBS at 4°C. 50-mm-thick coronal brain sections were cut with a microtome (HM650V, Microm, Walldorf, Germany) and stored at 4°C in PBS with 0.1% sodium azide.
After overnight post-fixation in PFA (4%) at 4°C, coronal brain sections (25 μm) were cut with a vibratome (VT1000E; Leica, Germany).
