α actin

Polyclonal rabbit IgG antibodies against human GAPDH, β-actin, phospho-/total-P38, phospho-/total-ERK1/2, phospho-/total-JNK, α-actin, collagen, phospho-/total-Akt, horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG, and anti-rabbit IgG antibodies were purchased from GeneTex (Irvine, CA, USA). Polyclonal rabbit antibodies against human collagen were purchased from Proteintech (Chicago, IL, USA). Monoclonal rabbit antibodies against human PCNA were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Monoclonal rabbit antibodies against human TGF-β and vimentin were purchased from Abcam (Cambridge, UK). Polyclonal rabbit IgG against human Iba-1 was purchased from Wako (Chuo-ku, Osaka, Japan). Polyclonal rabbit IgG against human MPO was purchased from MyBioSource (San Diego, CA, USA). MK2006, PD98058, SP600125, and SB203580 were purchased from Biomol (Plymouth Meeting, PA, USA). LPS and BrdU were purchased from Sigma-Aldrich (St. Louis, MO, USA). TRITC- and FITC-conjugated goat anti-mouse IgG antibodies were purchased from Jackson ImmunoResearch (West Grove, PA, USA).

For protein extraction, cells were washed twice with ice‐cold PBS and lysed on ice in golden lysis buffer (20 mmol/L Tris‐HCl (Sigma Aldrich RES3098T‐B7, St. Louis, MI), pH 8.0, 137 mmol/L NaCl (Sigma Aldrich S7653, St. Louis, MI), 5.95 mmol/L EDTA (Sigma Aldrich 1233508, St. Louis, MI), 5 mmol/L EGTA (Sigma Aldrich E3889, St. Louis, MI), 10 mmol/L NaF (Sigma Aldrich S7920, St. Louis, MI), 1% Triton X‐100 (Sigma Aldrich X100, St. Louis, MI), and 10% glycerol (Sigma Aldrich G5516, St. Louis, MI)) supplemented with protease inhibitors (Sigma Aldrich P8340, St. Louis, MI) and phosphatase inhibitors (Sigma Aldrich P2850, St. Louis, MI). The proteins were separated via 12% sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) and transferred to PVDF membranes (Thermo Fisher Scientific LC2002, CA). HAS3 (Sigma Aldrich SAB2108148, St. Louis, MI), GFAP (GeneTex GTX108711, CA), β‐tubulin (GeneTex GTX11307, CA), TUBB3 (GeneTex GTX631836, CA) and α‐actin (GeneTex GTX109639, CA) antibodies were diluted 1:2000 in TBST, and the membranes were incubated for 2 hours at room temperature. Horseradish peroxidase (HRP)‐conjugated anti‐mouse and anti‐rabbit IgG (Santa Cruz Biotechnology SC‐2005, CA) secondary antibodies were diluted 1:4000, and the membranes were incubated for 1 hour at room temperature. α‐Actin was used as the protein loading control.