Gene and protein expression in original and siRNA treated fibroblasts from TBR1CA; Cre-ER mice injected with 4-OHT. (A) Relative transcript levels of Col1a2, Ctgf, and SPARC in cultured fibroblasts transfected with non-targeting siRNA (NT siRNA), Ctgf siRNA and SPARC siRNA. The expression level of each gene in the fibroblast lines with NT siRNA transfection was normalized to 1. *, P < 0.05. (B) Western blot analysis of type I collagen (COL1), CTGF, and SPARC in the fibroblasts from constitutively active Tgfbr1 mice transfected with NT siRNA, Ctgf siRNA or SPARC siRNA. N, non-targeting siRNA transfected fibroblasts; C, Ctgf siRNA transfected fibroblasts; S, SPARC siRNA transfected fibroblasts. (C) Densitometric analysis of Western blots for protein level of COL1, CTGF, and SPARC. Compared to non-targeting siRNA treatment, Ctgf siRNA or SPARC siRNA transfected fibroblasts showed significant reduction of COL1 (P = 0.015 or 0.009 respectively). Significant reduction of CTGF (P = .002) by Ctgf siRNA and SPARC (P = 0.0004) by SPARC siRNA were also shown. Bars show the mean ± SD results of analysis of three independent experiments performed in triplicate. *, P < 0.05.
Wang et al. Arthritis Research & Therapy 2010 12:R60 doi:10.1186/ar2973