Relative expression of GFAP and Iba-1 by Western blotting

Total proteins were extracted from the CX and HP from both rCHIMERA and sham mouse perfused brains at 1 month after injury. The protein concentration was measured by BCA kit (life technologies; product# 1856136, CA, USA). The same amount of protein from each sample was separated on 4–12% Bis-Tris gels (Life technologies: NW04120BOX) under denaturing and reduced conditions, and transferred to nitrocellulose membranes. The membranes were probed with the following primary antibodies: GFAP (1:5000, Thermo Scientific: cat# MS-1376-P), Iba-1 (1:1000, Wako, 019-19741), phospho-tau (Ser202) (1: 1,000, Cell Signaling Technology, product# 11834, MA, USA) and GAPDH (1:5000, Cell Signaling Technology, product#: 5174S, MA, USA), and with peroxidase-conjugated F(ab’)² fragment goat anti-rabbit or anti-mouse IgG (H+L) as a secondary antibody (Jackson ImmunoResearch lab, Inc., code number: 111-036-003). The proteins were detected by Pierce supersignal west pico chemiluminescent detection substrate (Thermo Fisher Scientific, Rockford, IL, USA). All images of immunoblots were taken and quantified using KODAK 1D software (Eastman Kodak, NY, USA).