Figure 8

Gene expression in rib chondrocytes from Hoxc-8 and Hoxd-4 transgenic mice. Real-time quantitative PCR was performed as described. Panel A shows typical curves for real-time detection of amplification. Panel B shows a melting curve analysis for the PCR products, which is indicative of the quality of the reaction. For each reaction, the fidelity of amplification was assessed by inspection of the amplification and melting curves. Panel C lists the cycle number above the threshold level (Ct) for each primer pair at which product was detected in a representative experiment in FVB controls. Low Ct values reflect higher expression levels, high Ct values correspond to lower expression levels. Panel D shows gene expression measurements in Hoxc-8 transgenic samples. GAPDH cDNA levels in each sample were used to standardize measurements, and gene expression levels are normalized to the level of each gene found in primary chondrocytes from normal FVB control mice. The results are plotted as "fold change relative to FVB" with decreased expression in negative (left on X-axis) iand increased expression in positive values (right on X-axis). The FVB control sample would always be 1, and therefore was not plotted. RNA extracts were prepared from postnatal day 0 mouse rib chondrocytes from normal FVB newborns or animals transgenic for Hoxc-8, and equal amounts of cDNA were pooled from 4–5 individual animals for each sample.