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. 2021 Nov 10;10:e70992. doi: 10.7554/eLife.70992

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© 2021, Gastfriend et al

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Figure 1. — (A) Overview of the endothelial differentiation and Wnt treatment protocol. (B) Immunocytochemistry analysis of CD34 and CD31 expression in D5 endothelial progenitor cells (EPCs) prior to magnetic-activated cell sorting (MACS). Hoechst nuclear counterstain is overlaid in the merged image. Scale bars: 200 μm. (C) Flow cytometry analysis of CD34 and CD31 expression in D5 EPCs prior to MACS. (D) Immunocytochemistry analysis of β-catenin and GLUT-1 expression in Passage 1 ECs treated with Wnt3a or control. Hoechst nuclear counterstain is overlaid. Arrowheads indicate smooth muscle-like cells (SMLCs). Scale bars: 200 μm. (E) Quantification of the percentage of GLUT-1⁺ ECs in control- and Wnt3a-treated conditions. Points represent replicate wells from two independent differentiations of the IMR90-4 line, each differentiation indicated with a different color. Bars indicate mean values. p-value: two-way ANOVA.

Figure 1—figure supplement 1. — (A) Overview of the endothelial differentiation and Wnt treatment protocol. (B) Immunocytochemistry analysis of CD34 and CD31 expression in D5 endothelial progenitor cells (EPCs) prior to magnetic-activated cell sorting (MACS). Hoechst nuclear counterstain is overlaid in the merged image. Scale bars: 200 μm. (C) Flow cytometry analysis of CD34 and CD31 expression in D5 EPCs prior to MACS. (D) Immunocytochemistry analysis of β-catenin and GLUT-1 expression in Passage 1 ECs treated with Wnt3a or control. Hoechst nuclear counterstain is overlaid. Arrowheads indicate smooth muscle-like cells (SMLCs). Scale bars: 200 μm. (E) Quantification of the percentage of GLUT-1⁺ ECs in control- and Wnt3a-treated conditions. Points represent replicate wells from two independent differentiations of the IMR90-4 line, each differentiation indicated with a different color. Bars indicate mean values. p-value: two-way ANOVA.