Fig. 4.

qPCR validation of the microarray expression data. The relative mRNA expression levels of selected genes related to skeletal system development (a), ECM organization and bone formation (b), and GAG biosynthesis (c), were determined with the 2^−(ΔΔCt) method normalized with the geometric mean of the HPRT, ATP5B and CYC1 reference genes. Bars represent the mean ratio of target gene expression in patients' fibroblasts compared to three unrelated healthy individuals. qPCR was performed in triplicate, and the results are expressed as means ± SEM. In A and B, the relative mRNA levels of target genes in patients versus controls are expressed as Log10 transformed values. In C, the non-transformed relative fold change is shown. Statistical significance (*P < 0.05, **P < 0.01 and ***P < 0.001) was calculated with one-way ANOVA and the Tukey post hoc test.