Skip to main content
. 2013 Oct 3;11:75. doi: 10.1186/1478-811X-11-75

Figure 2.

Figure 2

Differential Smad7 expression leads to changes in the expression of CCN2 and type I collagen in CCD-1068SK fibroblasts. CCD-1068SK fibroblasts were transfected with 80 μM Smad7 siRNA and incubated for 48 hours before isolating RNA and protein for further analysis. Fibroblasts transfected with scrambled siRNA were used as a control. (A) Quantitative real-time RT-PCR results show levels of Smad7, CCN2, COL1A1 and COL1A2 mRNA relative to GAPDH (*p ≤ 0.05, ** p ≤ 0.01, n = 3). (B) Western blotting results show Smad7, α1(I) and α2(I) procollagen and CCN2 levels in Smad7 knock-down fibroblasts compared to control fibroblasts. The CCN2 antibody detected a band at 36-38 kDa and 72 kDa that represent monomeric and dimeric forms of the protein. (B) For Smad7 overexpression experiments, CCD-1068SK fibroblasts were transiently transfected with 1 μg of the pORF9-hSmad7 plasmid. RNA and protein was isolated 8 hours and 48 hours after transfection. (C) Relative levels of Smad7, CCN2, COL1A1 and COL1A2 were quantified by means of quantitative real-time RT-PCR analysis (*p ≤ 0.05, ** p ≤ 0.01, n = 3). (D) Antibodies against Smad7, CCN2 and type I collagen were used to detect the respective protein levels by means of Western blot analysis. β-tubulin was used as a loading control.