Western Blot Analysis of Brain Proteins

Western blot analysis was performed as previously described [38 (link)]. For details, see SI Methods. Briefly, mice were sacrificed by rapid decapitation under deep anesthesia. The brain tissues were dissected in ice-cold 1× PBS buffer, then the samples were homogenized and centrifuged. Total protein concentration was determined using an Enhanced BCA Protein Assay Kit (Beyotime). The following antibodies were used: rabbit anti-PV antibody (1:4000, Abcam), rabbit anti-Arg1 antibody (1:5000, Abcam), rabbit anti-β-tubulin antibody (1:5000, Cell Signaling Technology), rabbit anti-IL1β antibody (1:2000, Abcam), rabbit anti-IL6 antibody (1:3000, GeneTex), rabbit anti-TNFα antibody (1:3000, GeneTex), and goat anti-rabbit IgG (H + L) HRP (1:8000, Abways). Protein bands were visualized using a chemiluminescence system (ChemiDocTM XRS+, BioRad). The protein expressions were semi-quantitatively analyzed with ImageJ software.