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After a second wash, total RNA from pellets was isolated using the Qiagen [http://www.selleckchem.com/products/AZD0530.html Saracatinib chemical structure] MicroKit (Qiagen, Australia). Total RNA was also extracted from a control culture without the addition of any growth factor or inhibitor. Synthesis of cDNA and gene expression analysis of collagen-2a (with Cyclophilin-A as the internal reference) were conducted as described above (n?=?4 per treatment). Data are presented as mean?��?SEM. Comparisons between g?6976-treated and vehicle-treated groups were analyzed using one-way ANOVA. To explore the role of osterix during the mesenchymal differentiation phase of growth plate injury repair, g?6976 was used to inhibit the activation of protein kinase D which has been shown to potentially suppress transcriptional factor, osterix.24 Micro-CT analysis revealed that within the [http://www.selleckchem.com/products/CAL-101.html selleck chemicals] growth plate injury site, bone formation was observed in both injured groups, whilst non injured controls showed no signs of bone bridge formation within the growth plate area (Fig. 1A�CC). Analysis of bone volume present within the growth plate injury site showed that rats treated with the inhibitor resulted in a significant 22.26% reduction in total bone volume in comparison with those treated with vehicle (21.5?��?1.9 vs 27.7?��?2.0) (p?[http://en.wikipedia.org/wiki/SKAP1 SKAP1] mesenchymal tissue present within the injury site in comparison to rats of the vehicle group (30.9?��?4.6 vs 11.0?��?2.0) (p?