Fig. 4.

Suppression of CCN3/NOV by siRNA technology and fibrogenic gene expression in CFSC. a CFSC were transfected with an unspecific siRNA control, siRNAs (siNOV3, siNOV4, siNOV5, siNOV6) targeting CCN3/NOV expression, or an expression vector for rat NOV (rat NOV-1) and maintained in 0.5% FCS for 48 h. RNA was isolated from respective cells and subjected to qRT-PCR revealing that CCN3/NOV siRNA increased α-SMA, collagen type I and CCN2/CTGF mRNA expression, while CCN3/NOV overexpression showed no significant effects. b Quantitative RT-PCR showed that the transfection with siRNA targeting ccn3/nov gene expression resulted in decreased amounts of ccn3/nov mRNAs while the expression was significantly increased after transfection with expression vector rat NOV-1.c CCN3/NOV knock down efficacy was further demonstrated in ELISA testing using cell supernatants from the same experiment. d CFSC were transfected for 96 h with CCN3/NOV siRNAs and rat NOV-1 expression vector. Cell extracts were prepared and analysed by Western blot for expression of Fibronectin, collagen type I, α-SMA, CCN2/CTGF, TIMP-1 and β-Actin