transforming growth factor-beta 1 tgf-β1 10 ng/ml  (PeproTech)

Journal: Arthritis Research & Therapy

Article Title: Activation of proteinase-activated receptor 2 in human osteoarthritic cartilage upregulates catabolic and proinflammatory pathways capable of inducing cartilage degradation: a basic science study

doi: 10.1186/ar2329

Figure Lengend Snippet: Proteinase-activated receptor 2 (PAR-2) synthesis regulation. (a) PAR-2 immunostaining in osteoarthritis (OA) cartilage explants untreated ( n = 16) and treated with interleukin 1 beta (IL-1β) ( n = 9), tumor necrosis factor-alpha (TNF-α) ( n = 9), transforming growth factor-beta-1 (TGF-β1) ( n = 5), PAR-2-activating peptide (PAR-2-AP) 1 μM ( n = 3), PAR-2-AP 10 μM ( n = 4), PAR-2-AP 100 μM ( n = 4), and PAR-2-AP 400 μM ( n = 8) for 48 hours in Dulbecco's modified Eagle's medium (DMEM) 10% fetal calf serum (FCS). (b) Representative Western blot of PAR-2 synthesis in OA monolayer chondrocytes ( n = 3) incubated for 72 hours in DMEM 2.5% FCS in the absence (CTL) or presence of IL-1β, TGF-β1, PAR-2-AP 10 μM, PAR-2-AP 100 μM, and PAR-2-AP 400 μM. P values indicate the comparison with the untreated (CTL) specimens.

Article Snippet: Cartilage explants were treated for 48 hours by IL-1β (100 pg/mL), TNF-α (5 ng/mL), and transforming growth factor-beta-1 (TGF-β1) (10 ng/mL) (all from R&D Systems, Inc., Minneapolis, MN, USA) or by the synthetic PAR-2-activating peptide (PAR-2-AP), SLIGKV-NH 2 (0 to 400 μM) (Bachem California, Inc., Torrance, CA, USA), p38 inhibitor (SB 202190 at 10 μM) (Tocris Bioscience, Ellisville, MO, USA), and mitogen-activated protein (MAP) kinase kinase (MEK1/2) inhibitor (PD98059 at 10 μM) and nuclear factor-kappa B (NF-κB) inhibitor (SN50 at 50 μg/mL) (both from EMD Biosciences, Inc., San Diego, CA, USA).

Techniques: Immunostaining, Modification, Western Blot, Incubation, Comparison

Journal: Arthritis Research & Therapy

Article Title: Activation of proteinase-activated receptor 2 in human osteoarthritic cartilage upregulates catabolic and proinflammatory pathways capable of inducing cartilage degradation: a basic science study

doi: 10.1186/ar2329

Figure Lengend Snippet: Production of metalloproteinase (MMP)-1 (a) , MMP-13 (b) , and cyclooxygenase 2 (COX-2) (c) following interleukin 1 beta (IL-1β) and specific proteinase-activated receptor 2 (PAR-2) activation. Immunostaining data of osteoarthritic cartilage untreated ( n = 12) and treated with IL-1β ( n = 12), PAR-2-activating peptide (PAR-2-AP) 1 μM ( n = 3), PAR-2-AP 10 μM ( n = 4), PAR-2-AP 100 μM ( n = 4), and PAR-2-AP 400 μM ( n = 9). P values indicate the comparison with the untreated (CTL) specimens.

Article Snippet: Cartilage explants were treated for 48 hours by IL-1β (100 pg/mL), TNF-α (5 ng/mL), and transforming growth factor-beta-1 (TGF-β1) (10 ng/mL) (all from R&D Systems, Inc., Minneapolis, MN, USA) or by the synthetic PAR-2-activating peptide (PAR-2-AP), SLIGKV-NH 2 (0 to 400 μM) (Bachem California, Inc., Torrance, CA, USA), p38 inhibitor (SB 202190 at 10 μM) (Tocris Bioscience, Ellisville, MO, USA), and mitogen-activated protein (MAP) kinase kinase (MEK1/2) inhibitor (PD98059 at 10 μM) and nuclear factor-kappa B (NF-κB) inhibitor (SN50 at 50 μg/mL) (both from EMD Biosciences, Inc., San Diego, CA, USA).

Techniques: Activation Assay, Immunostaining, Comparison

Journal: Arthritis Research & Therapy

Article Title: Activation of proteinase-activated receptor 2 in human osteoarthritic cartilage upregulates catabolic and proinflammatory pathways capable of inducing cartilage degradation: a basic science study

doi: 10.1186/ar2329

Figure Lengend Snippet: Signalling pathways induced by specific proteinase-activated receptor 2 (PAR-2) activation. Representative Western blot of time and dose curves of PAR-2 signalling in osteoarthritic chondrocytes ( n = 3 or 4). Phosphorylated forms of Erk1/2 (p-Erk1/2) (a,b) , p38 (p-p38) (c,d) , and JNK (p-JNK) (e,f) treated with PAR-2-activating peptide (PAR-2-AP) at 0 (-), 10, 100, and 400 μM in the absence (a,c,e) or presence (b,d,f) of interleukin 1 beta (IL-1β) (100 pg/mL) for 0 to 60 minutes (a,c,e) . P values indicate the comparison between the untreated (-) and the PAR-2-AP-treated chondrocytes. For p-Erk1/2, all of the times and concentrations showed a statistically significant increase ( p < 0.03). For p-p38, statistical significance was reached for each concentration at 5 and 15 minutes ( p < 0.03). (b,d,f) P values indicate the comparison between the untreated and the IL-1β-treated chondrocytes in the absence or presence of PAR-2-AP. Erk1/2, extracellular signal-regulated kinase 1/2; JNK, c-jun N-terminal kinase.

Article Snippet: Cartilage explants were treated for 48 hours by IL-1β (100 pg/mL), TNF-α (5 ng/mL), and transforming growth factor-beta-1 (TGF-β1) (10 ng/mL) (all from R&D Systems, Inc., Minneapolis, MN, USA) or by the synthetic PAR-2-activating peptide (PAR-2-AP), SLIGKV-NH 2 (0 to 400 μM) (Bachem California, Inc., Torrance, CA, USA), p38 inhibitor (SB 202190 at 10 μM) (Tocris Bioscience, Ellisville, MO, USA), and mitogen-activated protein (MAP) kinase kinase (MEK1/2) inhibitor (PD98059 at 10 μM) and nuclear factor-kappa B (NF-κB) inhibitor (SN50 at 50 μg/mL) (both from EMD Biosciences, Inc., San Diego, CA, USA).

Techniques: Activation Assay, Western Blot, Comparison, Concentration Assay