Journal: Arthritis Research & Therapy
Article Title: Molecular targeting of hepatocyte growth factor by an antagonist, NK4, in the treatment of rheumatoid arthritis
Figure Lengend Snippet: AdCMV.NK4 inhibits inflammatory cell infiltration, as well as cytokine and RANKL expression, in the synovial tissues of SKG mice . Immunohistochemistry of interleukin 1 (IL-1) (A) , IL-6 (B) , TNF-α (C) , CD31 (D) , IL-17 (E) and RANKL (receptor activator of nuclear factor κB ligand) (F) . High-level expression of IL-1, IL-6, TNF-α, CD31, IL-17 and RANKL in the synovial tissues of LacZ-treated SKG mice was observed (upper). In contrast, the expression was significantly inhibited in the synovial tissues of NK4-treated SKG mice (lower). Original magnification, ×200.
Article Snippet: For immunohistochemical staining, cryostat sections of joints were fixed in cold acetone for 10 min, washed in PBS and depleted of endogenous peroxidase by treatment with 0.3% H2 O2 in absolute methanol for 15 min. After blocking nonspecific binding with 10% normal rabbit serum in PBS for 30 min, the sections were incubated with anti-mouse interleukin 6 antibody (IL-6 Ab; goat immunoglobulin G (IgG)), anti-mouse tumor necrosis factor α (TNF-α) Ab (goat IgG), anti-mouse CD31 Ab (goat IgG), anti-RANKL (anti-receptor activator of nuclear factor κB ligand) Ab (goat IgG), anti-mouse IL-17 Ab (goat IgG; all from Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti-mouse IL-1 Ab (goat IgG; R & D Systems, Minneapolis, MN, USA), anti-mouse HGF Ab (rabbit IgG) or anti-mouse c-Met Ab (rabbit IgG) at appropriate dilutions for 1 h at room temperature, washed, incubated with biotinylated goat anti-rabbit IgG, washed again and incubated with avidin-biotinylated horseradish peroxidase complex (ABC) and 3,3'-diaminobenzidine tetrahydrochloride (DAB) (VECTASTAIN Elite ABC Kit; Vector Laboratories, Burlingame, CA, USA) and counterstained with Mayer's hematoxylin.
Techniques: Expressing, Mouse Assay, Immunohistochemistry