Quantitative Western Blot Analysis of Neuroinflammatory Markers

Protein from each right hemisphere sample was extracted, quantified, and denatured at 95°C in 5X loading buffer for 10 min. Western blot was performed as previously described (17 (link)). The following primary antibodies were used: Fibrin (ogen) (ab34269, Abcam), P-selectin (60322-1-Ig, Proteintech), PDGFRβ (ab69506, Abcam), CypA (ab41684, Abcam), N-cadherin (22018-1-AP, Proteintech), phospho-NF-κB p65 subunit (p-p65, ab86299, Abcam), MMP-9 (10375-2-AP, Proteintech), and β-actin (66009-1-Ig, Proteintech). The bands were visualized using a BeyoECL Plus kit (P0018; Beyotime) and photographed by a chemiluminescence imaging system (ChemiDoc XRS+; Bio-Rad, Hercules, CA, USA). Band densities were quantified by a blinded observer using ImageJ software.