Fig. 4

Quantitative RT-PCR of transplanted forepaw tissues shows persistently increased expression of fibrosis genes in DD-cells versus CT-cells. Real time qRT-PCR was performed to measure the mRNA expression levels of α-SMA (a), type I collagen (b) and type III collagen (c), all of which were markedly elevated in DD-cells compared to CT-cells. For quantitative analysis of gene expression, the comparative threshold cycle (Ct) method for relative quantification was used. The expression of the target genes was normalized to GAPDH expression. Values represent the mean ± SEM of two separate experiments performed on RNA derived from the forepaw tissues of four animals, each performed in triplicate. Parallel in vitro cultures of the CT- and DD-derived fibroblasts transplanted in vivo were used as controls. P value less than 0.05 was considered as statistically significant