Connective tissue growth factor dependent collagen gene expression induced by MAS agonist AR234960 in human cardiac fibroblasts
Fig 1
Upregulation of CTGF, MAS and collagen in human heart failure.
(A) Bar graphs showing real-time PCR analysis of fold increase (2‒ΔΔCt) of MAS receptor mRNA (left) and CTGF mRNA (right) expression in tissue from failed human heart (HF) compared with non-failing (NF) samples (**p<0.01; ***p<0.001). Expression was normalized to GAPDH. (B) Correlation plot between normalized expression of CTGF and MAS mRNA showing strong interaction between them (p value–0.174). (C) Western-blot showing significant upregulation of CTGF in HF tissue samples compared to NF samples (right); GAPDH was used as loading control. The western blot image shown is representative of all the experiments done under similar experimental conditions and the data from multiple experiments quantitated and cumulative data were presented as bar graphs (left) (***p<0.001). (D) Masson’s Trichrome staining of cryo-sections (4μm) of human heart left ventricular wall tissue showing increased collagen deposition (stained blue) in inter-cellular spaces (arrows) of the HF samples compared to NF samples (upper panels); magnification ×20. Immuno-histochemical staining of same set of tissue sections with CTGF antibody, showing more deposition of CTGF (intense brown) in HF sections then NF samples (lower panels); magnification ×20. (E) Bar graphs showing real-time PCR analysis of different sub-types of collagen expression (Col 1A1, Col 1A2, Col 3A1 and Col 4A2) [represented as fold increase (2‒ΔΔCt)] in left ventricular heart tissue from failing (HF) as well as non-failing (NF) samples (***p<0.001). Expression was normalized to GAPDH. All the bar graphs are presented with error bar of ±SD.