Journal: Biochimica et biophysica acta

Article Title: Human T-cell leukemia virus type 1 tax transactivates the matrix metalloproteinase 7 gene via JunD/AP-1 signaling.

doi: 10.1016/j.bbamcr.2011.02.002

Figure Lengend Snippet: Fig. 10. Neutralization of MMP-7 using an MMP-7 polyclonal antibody (A) and JunD knockdown (B) inhibited SLB-1 cell invasion. (A) SLB-1 cells were seeded with or without a goat polyclonal MMP-7 neutralizing antibody (top and bottom panels) or with normal goat IgG (bottom panel) in the upper chamber for 24 h. (B) SLB-1 cells were transfected with either JunD or control siRNA and then incubated for 24 h. Thereafter, SLB-1 cells were seeded in the upper chamber for 24 h. Cell invasion was monitored using RT-CES system for SLB-1 cells. Data are mean±SD of three independent invasion experiments in a period of 24 h. *Pb0.05 vs. control.

Article Snippet: Rabbit polyclonal antibodies to JunD, phospho-c-Jun (Ser73), phospho-JunD (Ser100), c-Jun NH2-terminal kinase (JNK) and phospho-JNK (Thr183 and Tyr185), and rabbit monoclonal antibodies to extracellular signal-regulated kinase (ERK)1/2 and phospho-ERK1/2 (Thr202 and Tyr204) were purchased from Cell Signaling Technology (Beverly, MA, USA).

Techniques: Neutralization, Knockdown, Transfection, Control, Incubation

Journal: Arthritis Research & Therapy

Article Title: Hyaluronic acid fragments enhance the inflammatory and catabolic response in human intervertebral disc cells through modulation of toll-like receptor 2 signalling pathways

doi: 10.1186/ar4274

Figure Lengend Snippet: The effect of gene silencing on fHA-mediated IL-6 production in IVD cells . A ) siRNA-mediated knockdown of genes Toll like receptor ( TLR ) 2 , TLR4 , CD44 and RHAMM was confirmed after 30 hours in intervertebral disc (IVD) cells by qRT-PCR ( n = 4). In each case, gene expression was calculated as fold change as compared to untreated cells. The use of a non-specific scrambled siRNA (siRNA (S)), confirmed specificity of gene knockdown. B, C ) Interleukin (IL)-6 protein production by Pam3CSK4- (25 ng/ml) ( n = 4) ( B ) and lipopolysaccharide (LPS)- (25 ng/ml) ( n = 3). ( C ) stimulated IVD cells following gene knockdown of TLR2 or TLR4 respectively, as determined by IL-6 ELISA. ( D ) hyaluronic acid fragment (fHA)-treated (20 μg/ml) IVD cells following gene knockdown as determined by IL-6 ELISA ( n = 4). IL-6 protein levels are represented as a percentage of those measured for untreated cells. In all cases, analyses were performed in triplicate and values expressed as mean ± S.D. Statistical analysis was performed using the Student's t -test, * P <0.01.

Article Snippet: Cells were then pre-incubated for one hour with either an affinity purified polyclonal rat anti-human TLR2 neutralizing antibody (final concentration 5 μg/ml) (LabForce, Switzerland) or an isotype matched IgG control (Lucerna-Chem, Luzern, Switzerland).

Techniques: Knockdown, Quantitative RT-PCR, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: Arthritis Research & Therapy

Article Title: Hyaluronic acid fragments enhance the inflammatory and catabolic response in human intervertebral disc cells through modulation of toll-like receptor 2 signalling pathways

doi: 10.1186/ar4274

Figure Lengend Snippet: The effect of TLR2 inhibition on fHA-mediated IL-6 production in IVD cells . Interleukin (IL)-6 protein production by Pam3CSK4- (25 ng/ml) ( A ) and hyaluronic acid fragment (fHA)-treated (20 μg/ml) ( B ) intervertebral disc (IVD) cells following antibody-mediated neutralization of Toll like receptor (TLR)2 activity ( n = 4). IL-6 protein levels are represented as a percentage of those measured for untreated cells. In all cases, analyses were performed in triplicate and values expressed as mean ± S.D. Statistical analysis was performed using the Student's t -test, * P <0.05 as compared to cells treated with non-specific IgG control antibody.

Article Snippet: Cells were then pre-incubated for one hour with either an affinity purified polyclonal rat anti-human TLR2 neutralizing antibody (final concentration 5 μg/ml) (LabForce, Switzerland) or an isotype matched IgG control (Lucerna-Chem, Luzern, Switzerland).

Techniques: Inhibition, Neutralization, Activity Assay, Control