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. 2022 Jul 26;13(7):650. doi: 10.1038/s41419-022-05064-5

Fig. 1. The effect of ECs culture media on VSMCs calcification.

Fig. 1

A–C 5% calf serum (CS), 5% healthy serum (HS), and 5% uremia serum (US) were supplemented to the culture media (CM) of mice ECs, respectively. These CM were collected for the following VSMCs cultures. Alizarin Red S staining (A), ALP activity (B), and Runx2 expression (C) were measured on day 14, day 7, and day 7, respectively. The black arrows indicate mineralized nodules in VSMCs. D–F VSMCs were cultured with 0.9 mM inorganic-induced endothelial CM (ECsNPi-CM), 3.5 mM inorganic phosphate-induced endothelial cells CM (ECsHPi-CM), and their EV-depleted conditional media (ECsHPi-CM-EVs, ECsHPi-CM-EVs, and ECsHPi-CM-EVs + GW4869). Alizarin Red S staining (D), ALP activity (E), and Runx2 expression (F) were measured on day 14, day 7, and day 7, respectively (Scale bar = 100 μm). Three independent experiments were performed, and representative data were shown. Data are shown as mean ± SD. ns: no significance, *p < 0.05, **p < 0.01.