Fig. 3
Exosomes derived from AFs induced by high phosphorus promote VSMCs calcification. A Western blot was used to determine the effect of AFsNPi-Exos or AFsHPi-Exos treatment on Runx2 and BMP2 protein levels in 3.5 mM Pi-induced VSMCs. VSMCs treated with 0.9 mM Pi plus PBS were used as control. B Representative images and quantitative analysis of Alizarin red staining in 3.5 mM Pi-induced VSMCs cultured with AFsNPi-Exos or AFsHPi-Exos. The black arrows indicate mineralized nodules in VSMCs. The scale bar represents 500 μm. ALP staining (C) and ALP activity (D) of AFsNPi-Exos or AFsHPi-Exos treated VSMCs under 3.5 mM Pi treatment. Representative micrographs are shown. The scale bar represents 500 μm. E The transwell co-culture system was used to detect the calcification of VSMCs co-cultured with AFs pretreated with or without GW4869 under the treatment of NPi or HPi for 3 days. Western blot analysis of Runx2 and BMP2 protein levels (F) and ALP activity (G) in VSMCs co-cultured with AFs. Data are presented as mean ± SD from triplicate experiments. *p < 0.05, **p < 0.01