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. 2024 Mar 8;13(6):481. doi: 10.3390/cells13060481

Figure 1.

Figure 1

Indoxyl sulfate (IS) induces VEC osteogenic changes. Human valve endothelial cells (VEC) were treated with IS or PBS for 3, 5, and 7 days and harvested for cell viability, mRNA expression, and protein expression studies. (A) VECs were treated with IS (250 μM and 100 μM) or PBS and studied for cell viability using an MTT assay. Assays were performed in triplicates (*** p < 0.0001; n = 3); (B) RT-qPCR of CT- and IS-treated hVEC extracts showing mRNA expression of RUNX2 (* p < 0.01; n = 4); (C) Western blot analysis and quantification of RUNX2 in hVECs. Experiments were performed in duplicate (* p < 0.01; n = 3); (D) RT-qPCR of CT- and IS-treated hVEC extracts showing mRNA expression of alkaline phosphatase (ALP) (** p < 0.001; *** p < 0.0001; n = 3); (E) hVECs cultured with pro-osteogenic medium for 15 days were treated with IS (250 μM) or with PBS. Scale bar = 100 μm. Right panel: Quantification of alizarin-red positive area. n = 6. ** p < 0.001 vs. CT.