Array System, used to measure relative expression of 16 common endogenous control genes. Data analysis was performed using ABI’s Sequence Detection Systems (SDS) software 2.1, and the gene showing the least deviation in Ct values across all samples (β-Tubulin, B2M) was selected as the endogenous control gene for subsequent RT-PCR validation experiments. Validation was performed (n = 8/group) using the 7500 Fast Real-Time RT-PCR Gene Expression System (ABI). Each cDNA (20 ng) sample processed as above was amplified in triplicate using gene specific (β3-integrin, Lrp5, Ctnnb1, Dkk1, OPG) and control (B2M) TaqMan Gene Expression Assay primer/probes sets. Data was analyzed with SDS software 1.4 using the 2−ΔΔCT relative quantification method (Livak and Schmittgen, 2001).
