Research Paper Volume 12, Issue 21 pp 21220—21235

Cyclooxygenase-2/sclerostin mediates TGF-β1-induced calcification in vascular smooth muscle cells and rats undergoing renal failure

Figure 2. Effects of TGF-β1 on high phosphate-induced calcification markers in VSMCs cells. (A) Real-time PCR assay results show the effect of high phosphate and/or LY364947 on mRNA of Runx2 in VSMCs cells ("*" p<0.05 vs. control; "#" p<0.05 and "##” p<0.01 vs. high phosphate group). (B) Real-time PCR assay results show the effect of high phosphate and/or LY364947 on mRNA expression of SM22α in VSMCs cells ("*" p<0.05 and "**" p<0.01 vs. control; "#" p<0.05 vs. high phosphate group). (C) Western blot assay results show the effect of high phosphate and/or LY364947 on Runx2 and α-SMA in VSMCs cells; GAPDH was used as a loading control. (D) Quantification results of Western blot assay show the effect of high phosphate and/or LY364947 on Runx2 in VSMCs cells ("**" p<0.01 vs. control; "#" p<0.05 and "##" p<0.01 vs. high phosphate group). (E) Quantification results of Western blot assay show the effect of high phosphate and/or LY364947 on α-SMA in VSMCs cells ("**" p<0.01 vs. control; "#" p<0.05 and "##" p<0.01 vs. high phosphate group). (F) Western blot assay results show the effect of high phosphate and/or LY364947 on OPN in VSMCs cells; GAPDH was used as a loading control. (G) Quantification results of Western blot assay show the effect of high phosphate and/or LY364947 on OPN in VSMCs cells ("**" p<0.01 vs. control; "#" p<0.05 vs. high phosphate group). (H) Quantitative analysis results of Alizarin Red S staining show the effect of TGF-β1 on mineralization in VSMCs ("**" p<0.01 vs. control; "##" p<0.01 vs. high phosphate group). Pi: phosphate, LY364947: TGF-β type I receptor (TGFβRI) specific inhibitor.