Ma5 17 139

CMCS (MW 20 − 30 kDa, degree of carboxylation ∼80%, Beijing Solarbio Science & Technology Co., Ltd., CHN), TA (J&K Scientific Ltd., CHN), Cy5 (RuixiBio, CHN), phosphate buffered saline (PBS, Solarbio, CHN), normal saline (0.9%, China Resources Double-crane Pharmaceutical Co., Ltd., CHN), paraformaldehyde (PFA, 4%, Solarbio, CHN), sodium pentobarbital (Merck, Germany), and rose bengal (Sigma-Aldrich, Germany) were used.
Primary antibodies for IF staining:
Rabbit anti-IBA-1 (ab178846, Abcam, UK), mouse anti-iNOS (MA5-17,139, Invitrogen), goat anti-CD206 (AF2535, R&D), mouse anti-PSD95 (ab2723, Abcam, UK), and rabbit anti-Vglut1 (GTX133148, GeneTex, USA) were used.
Primary antibodies for WB:
Rabbit anti-CD206 (24,595, CST, USA), rabbit anti-iNOS (GB113965, Solarbio, CHN), mouse anti-p65 (GB12142, Solarbio, CHN), rabbit anti-p-p65 (GB113882, Solarbio, CHN), rabbit anti-IĸBα (GB111509, Solarbio, CHN), rabbit anti-p-IĸBα (AF2002, AFFinity, USA), rabbit anti-Synaptophysin (GB11814, Solarbio, CHN), rabbit anti-PSD95 (GB11277, Solarbio, CHN), rabbit anti-CD16 (80,006, CST, USA), rabbit anti-IL-1β (GB11113, Solarbio, CHN), rabbit anti-TGF-β (GB111876, Solarbio, CHN), rabbit anti-TLR4 (GB11519, Solarbio, CHN), rabbit anti-LaminB1 (GB111802, Solarbio, CHN), and mouse anti-GAPDH (GB15002, Solarbio, CHN) were used.

Four-μm-thick sections of the formalin-fixed-paraffin-embedded tissue samples were deparaffinized, incubated with 3% H₂O₂ for 10 min, underwent heat induced epitope retrieval (pH 6.0 sodium citrate buffer, 30 min in 80 °C water bath), additionally blocked with Background Block (Cell Marque, Rocklin, CA, USA) and incubated separately with mouse monoclonal primary antibodies against α-smooth muscle actin (α-SMA) (A2547, Merck, US, diluted 1:400), inducible nitric oxide synthase (iNOS) (MA5-17139, Invitrogen, US, diluted 1:400), or arginase-1 (Arg1) (ab239731, Abcam, UK, diluted 1:200) and detected by HRP-conjugated secondary goat antibodies (G-21040, Invitrogen, US, diluted 1:1000) and diaminobenzidine (DAB) with hematoxylin counterstaining.