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. 2022 Nov 10;13:1037230. doi: 10.3389/fphys.2022.1037230

FIGURE 3.

FIGURE 3

Activation of Piezo1 by Yoda1 increases Pi-mediated osteogenic differentiation and extracellular matrix (ECM) calcification of HAoSMCs and aorta rings. (A–C) HAoSMCs were cultured in osteogenic medium (OM; excess Pi: 1.5 mmol/L, excess calcium: 0.6 mmol/L) in the presence or absence of Yoda1 (10 μmol/L). (A) Representative Alizarin Red staining (day 6) and quantification. (B) Calcium accumulation in the ECM (day 6). (C) Osteocalcin (OCN) level (day 6) in EDTA-solubilized ECM samples. (D–E) Aortic rings obtained from C57BL/6 mice were cultured in control, OM and OM + Yoda1 (OMY) conditions. (D) Calcium content of aorta rings normalized to protein level (day 6). (E–F) Histological analysis of ex vivo cultured aortic rings from C57BL/6 mice. (E) Representative Hematoxylin-eosin (H&E) and von Kossa–stained aortic sections of untreated, OM, and OM + Yoda1-treated (OMY) aorta rings (day 7). Magnification, ×100, ×400. (F) Quantification of von Kossa staining. Data are shown as mean ± SD, n = 3–8. *p < 0.05, **p < 0.01, ***p < 0.005, ****p < 0.001.