Investigating Inflammatory Signaling Pathways

Recombinant human TNF-α was purchased from Peprotech and 2′,7′-dichlorofluorescein diacetate (DCF-DA) was purchased from Sigma–Aldrich. The following antibodies were used: anti-SOD3 for western blotting (ab83108) and anti-SOD3 for immunohistochemistry (IHC; ab171738) from Abcam; anti-glyceraldehyde dehydrogenase (GAPDH; ab83108) from AbClone; anti-phospho-NF-κB inhibitor protein ɑ (IκBɑ), anti-phospho-NF-κB p65, anti-phospho-c-jun N-terminal kinase (JNK), anti-JNK, anti-phospho-p38 mitogen-activated protein kinase (MAPK) (Cell Signaling Technology), anti-NF-κB p65, anti-p38 MAPK, anti-phospho-extracellular signal-regulated kinase (ERK), and anti-ERK2 from Santa Cruz Biotechnology; and anti-collagen type I cleavage site from ImmunoGlobe. Anti-MMP-1 and procollagen α1(I) N-propeptide (pN-ColIα1) antibodies have been previously described [21 (link)]. Horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were purchased from KOMA Biotech (Seoul, Korea). Alexa Fluor^® 488 goat anti-rabbit IgG (H+L) and rhodamine Red-X-conjugated goat anti-mouse IgG were purchased from Thermo Fisher Scientific (Waltham, MA, USA). SPlink HRP Detection Bulk Kit for Mouse and Rabbit Antibodies and Liquid AEC Substrate Kit (20×) for IHC were purchased from GBI labs (Bothell, WA, USA).