Fig. 1
The conditioned medium of HPi-induced AFs promotes VSMCs calcification. VSMCs were treated with 0.9 mM or 3.5 mM inorganic phosphate-induced AFs conditioned medium (AFsNPi-CM, AFsHPi-CM), 0.9 mM or 3.5 mM inorganic phosphate-induced AFs exosomes-free conditioned medium (AFsNPi-CM− Exos, AFsHPi-CM− Exos), 3.5 mM inorganic phosphate-induced GW4869-pretreated AFs conditioned medium (AFsHPi-CM + GW4869). A Western blot and quantitative analysis of Runx2 and BMP2 protein levels in VSMCs. B The formation of mineralized nodules in VSMCs was detected by Alizarin red staining. The black arrows indicate mineralized nodules in VSMCs. Representative microscopic views are shown. The scale bar represents 500 μm. C ALP activity in VSMCs was measured by the ALP kit. D ALP staining and quantitative analysis of the percentage of ALP staining positive cells in total area. Representative microscopic views are shown. The scale bar represents 500 μm. Three independent experiments were performed, and representative data are shown. Data are shown as mean ± SD. *p < 0.05, **p < 0.01