Expression of cyclooxygenases during chondrogenic differentiation from progenitor cells.

<p><b>A:</b> Expression of COX-1 and COX-2 in growth plates was determined by IHC. RZ = resting zone, PZ = proliferative zone, HZ = hypertrophic zone. <b>B:</b> Col2a1, Col10a1, Sox9 and Runx2 mRNA expression during chondrogenic differentiation of ATDC5 cells. <b>C:</b> COX-1 and COX-2 mRNA expression. In graphs, error bars represent mean ± s.e.m.(standard error of the mean; n = 3). * indicates <i>p</i> < 0.05. <b>D:</b> Protein expression of COX-1 and COX-2 in differentiation. Molecular weight markers (kDa) are depicted on the left of immunoblots and relative quantifications are depicted on top of immunoblots. <b>E:</b> Medium concentrations of PGD₂, PGE₂, PGF_2α and TXA₂ during chondrogenic differentiation.</p

COX-1 and COX-2 specific inhibitors differentially influence prostaglandin levels during ATDC5 chondrogenic differentiation.

<p>Successful COX-1 or COX-2 inhibition by the inhibitors and possible differences in specific prostaglandin production was determined at day 14 in ATDC5 differentiation. NS398, Celecoxib, SC-50 or Mofezolac were added from the start of differentiation or from day 10 onwards. <b>A:</b> PGE₂, PGD₂, PGF_2α and TXA₂ medium concentrations were determined at day 14 in differentiation by prostaglandin specific ELISA. <b>B:</b> COX-1 and COX-2 mRNA expression at day 14 in differentiation was determined by RT-qPCR (relative to t = 0 and corrected for β-actin). In graphs, error bars represent mean ± s.e.m. (n = 3). * indicates <i>p</i> < 0.05.</p

Effects of COX-1 and COX-2 specific inhibitors on chondrogenic differentiation of ATDC5 cells.

<p><b>A:</b>. Col2a1, Col10a1, Sox9 and Runx2 mRNA expression was determined at day 14 in ATDC5 differentiation. <b>B:</b> Protein expression of Col2a1 and Col10a1 at day 14 in differentiation. Molecular weight markers (kDa) are depicted on the left of immunoblots and relative quantifications are depicted on top of immunoblots. <b>C:</b> Glycosaminoglycans (GAGs) were stained and fold change of t = 14 samples was calculated as compared to t = 0 samples. Fold change (DNA) from samples from t = 14 was calculated relatively to day 0. In graphs, error bars represent mean ±s.e.m. (n = 3). * indicates <i>p</i> < 0.05.</p

Primer sequences for RT-qPCR.

<p>Primer sequences for RT-qPCR.</p