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. 2020 Aug 27;21(17):6210. doi: 10.3390/ijms21176210

Figure 5.

Figure 5

Natural and artificial P2 receptor agonists and antagonists influenced hMSC differentiation towards endothelial cells and smooth muscle cells. Endothelial and smooth muscle cell differentiated cells were evaluated by tube formation assay, eNOS and calponin staining separately. (A) Both ATP and UDP-glucose can significantly enhance tube formation. Scale bar 200 µm; (B) ATP and ADP decreased calponin expression while PPADS and suramin reversed these effects. Scale bar 100 µm; (C,F) P2X7 artificial antagonist A74003 and P2Y1 antagonist MRS2500 enhanced the number of calponin-positive cells, P2Y1 agonist MRS2365 nearly inhibited smooth muscle cell differentiation, while P2X3 antagonist RO-3 seemed to have no effect. Scale bar 100 µm; (D) Endogenous nucleotide released during the endothelial cell and smooth muscle cell differentiation increased tube formation Scale bar 200 µm; (E) while inhibiting smooth muscle cell differentiation. Scale bar 200 µm; (F) Quantification of endothelial and smooth muscle cell differentiation and treatment with natural and artificial agonist and antagonists. ** p < 0.05; *** p < 0.001. (Immunofluorescent pictures see Supplementary Materials Figures S4 and S5).