Journal: Journal of molecular medicine (Berlin, Germany)

Article Title: Genetic ablation of adenosine receptor A3 results in articular cartilage degeneration

doi: 10.1007/s00109-018-1680-3

Figure Lengend Snippet: A3 receptor agonist prevents articular cartilage matrix degradation induced by TNF-alpha and hypo-osmotic shock. Collagenase-generated collagen neo-epitope (a) and aggrecanase-generated aggrecan neo-epitope (b) in the media of pig articular cartilage explants as visualized by Western blot. Explants were cultured for 24 h in the indicated conditions. A3 receptor agonist, IB MECA (1 nM), was able to decrease collagenase and aggrecanase activity in culture as shown by the downregulation of aggrecan neo-epitope and collagen type II neoepitope expression. Data shown are normalized to explant wet weight. Data represent four biological replicates and are presented as mean ± SD; P values were calculated with one-way ANOVA followed by Tukey honest significant difference post hoc test. P values < 0.05 were considered to be significant

Article Snippet: Proteins were separated and analyzed by Western blot as mentioned above using aggrecan neo-epitope (Novus Biologicals, Littleton, CO) and collagen neo-epitope primary antibodies (IBEX Pharmaceuticals Inc. Mount Royal, QC).

Techniques: Generated, Western Blot, Cell Culture, Activity Assay, Expressing

Journal: PLoS ONE

Article Title: CCN Family Member 2/Connective Tissue Growth Factor (CCN2/CTGF) Has Anti-Aging Effects That Protect Articular Cartilage from Age-Related Degenerative Changes

doi: 10.1371/journal.pone.0071156

Figure Lengend Snippet: (A) Toluidine blue staining of frontal sections of medial portion of TG (a, c, e) and WT (b, d, f) littermate knee joints from 5- (a and b), 12- (c and d), and 24- (e and f) month-old male mice. TB staining indicated that WT articular cartilage was degraded with age, and finally the whole layer was lost after 24 months. (B) Immunohistochemical staining of aggrecan neoepitope in frontal sections of the medial portion of TG (a, c, e) and WT (b, d, f) littermate knee joints from 5- (a and b), 12- (c and d), and 24-month-old (e and f) mice. a’, b’, c’, d’, e’, and f’: higher magnification of the medial tibial plateau in the load-bearing region, indicated by the dotted box in a, b, c, d, e, and f, respectively. Bars: 200 µm. In the WT tibia (d and d’) even the remaining matrices were slightly degraded, as shown by the aggrecan neoepitope staining. TG cartilage showed a minor decrease in staining for the aggrecan neoepitope (a’, c’, and e’), but not severe as in the WT cartilage (b’, d’, and f’).

Article Snippet: Sections of knee joints were treated with bovine testicular hyaluronidase (25 mg/ml) for 30 minutes at room temperature for epitope retrieval and then immunostained with anti-CCN2 (Abcam, Cambridge, UK), anti-aggrecan neoepitope (Novus Biologicals, Littleton, CO), anti-type I (Millipore, Billerica, MA), type II (CII D3, ), and type X collagen (X53, both kindly provided by Dr. von der Mark, University of Erlangen) and anti-MMP13 antibodies (Millipore).

Techniques: Staining, Immunohistochemical staining