Journal: Arthritis Research & Therapy

Article Title: Connective tissue growth factor promotes articular damage by increased osteoclastogenesis in patients with rheumatoid arthritis

doi: 10.1186/ar2863

Figure Lengend Snippet: Connective tissue growth factor mediate ERK1/2 and focal adhesion kinase activation through integrin αVβ3 signal transduction. Figure 6A shows the immnoprecipitation and immunoblotting analysis. The cell extracts of osteoclasts stimulated with recombinant CTGF (10 or 50 ng/ml) at 60 minutes were precipitated using anti-integrin αVβ3 antibody and subsequently blotted with anti- phosphorylated ERK1/ERK2, conventional ERK1/ERK2, and integrin αVβ3 antibodies respectively. The phosphorylated ERK1/ERK2 was recruited with integrin αVβ3 by CTGF stimulation. Figure 6B shows the immunoblotting analysis using anti- phosphorylated FAK, conventional FAK, and β-actin antibodies in the osteoclasts extracts treated with CTGF (10 ng/ml) at 5, 15, 60, and 120 minutes in the presence or absence of anti-CTGF antibody (1 μg/ml). CTGF stimulation resulted in phosphorylation of FAK from 60 minutes and this effect was neutralized by anti-CTGF antibody suggesting activation of signal transduction pathways through integrin αVβ3.

Article Snippet: The primary antibodies; mouse anti-human β actin antibody (Sigma) at 1:500 dilution, rabbit anti-human focal adhesion kinase (FAK) antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) at 1:200 dilution, rabbit anti-human phospholilated FAK at 1:200 dilution were incubated with the blots at 37°C, 120 minutes.

Techniques: Activation Assay, Transduction, Western Blot, Recombinant

Journal: Molecular Medicine Reports

Article Title: SCRG1 suppresses LPS-induced CCL22 production through ERK1/2 activation in mouse macrophage Raw264.7 cells

doi: 10.3892/mmr.2017.6492

Figure Lengend Snippet: Several roles of scrapie responsive gene 1 (SCRG1) suggested in this study and our previous reports. SCRG1 is hypothesized to be secreted into the extracellular space by mesenchymal sem cells (MSCs) and to have autocrine/paracrine activity. The putative receptor for SCRG1 is a complex of bone marrow stromal cell antigen 1 (BST1) and β1 or β2 integrins on the cell surface. The SCRG1/BST1 axis positively regulates the self-renewal, migration, and osteogenic differentiation potentials of MSCs through the FAK/PI3 K/Akt signaling pathway. In macrophage, lipopolysaccharide (LPS) interacts with a heterologous receptor involving toll-like receptor 4 (TLR4), CD14, and MD2. SCRG1 suppresses LPS-induced CC-chemokine ligand 22 (CCL22) production of macrophage in a extracellular signal-regulated kinase 1/2 (ERK1/2)-dependent manner.

Article Snippet: The following primary antibodies that has been purchased from Cell Signaling Technology (Danvers, MA, USA) were used; anti-p44/42 mitogen-activated protein kinase (ERK1/2), anti-phospho-ERK1/2, anti-c-Jun N-terminal kinase (JNK), anti-phospho-JNK, anti-p38, anti-phospho-p38, anti-Akt, anti-phospho-Akt, anti-focal adhesion kinase (FAK), anti-phospho-FAK. β-actin level measured were detected with an anti-β-actin antibody (Santa Cruz Biotechnology, Dallas, TX, USA) as a loading control.

Techniques: Activity Assay, Migration