IL-21 promotes osteoblastic differentiation of human valvular interstitial cells through the JAK3/STAT3 pathway

Figures

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  Figure 1. IL-21 expression in human calcific AVs. A) Relative mRNA expression. B-E) IL-21, ALP, and Runx2 expression in human calcific AVs. F) Representative images of immunohistochemical staining for IL-21 in (top row, 4X) normal and (bottom row, 8X) calcific AVs; n = 5 for each group. **P < 0.01.

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  Figure 2. IL-21 protein induces osteoblastic differentiation of aortic valve interstitial cells. A-C) ALP and Runx2 expression in VICs (n = 3). D-F) Time course of IL-21 induction of ALP and Runx2 expression in VICs (n = 3). G) Effect of IL-21 (50 ng/ml) treatment on ALP and Runx2 mRNA expression in VICs for 3 days (n = 3). H-I) IL-21 induces deposition of calcium in VICs (n = 3). NS >0.05, *P < 0.05, **P < 0.01, ***P < 0.001.

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  Figure 3. Effect of JAK3 inhibition on osteogenic differentiation of VICs induced by IL-21. A-C) Expression of Runx2 and ALP in VICs treated with IL-21 for 3 days, then treated with WHI-P154. D-E) ALP and Runx2 mRNA expression. F) Alizarin Red S staining was used to observe calcium deposition in human AVICs after 21 days in culture with different interventions, NS >0.05, *P < 0.05, **P < 0.01, ***P < 0.001.

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  Figure 4. STAT3 knockdown inhibits IL-21-induced osteoblastic differentiation of VICs. A) Expression of STAT3 48 h after siRNA introduction. B-D) Western blotting showing inhibition of IL-21-induced expression of ALP and Runx2. E-F) ALP and Runx2 mRNA expression. G) Alizarin Red S staining was used to observe calcium deposition in VICs after 21 days in culture with different interventions. NS >0.05, *P < 0.05, **P < 0.01, ***P < 0.001.

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  Figure 5. Schematic of IL-21 promotion of osteoblastic differentiation of VICs through the JAK3-STAT3 pathway.