Figure 7.
Figure 7. Isl1 and Mesp1 cooperate in promoting cardiovascular differentiation in ESCs. (A) Schematic representation of Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs. (B) FACS analysis of CXCR4, PDGFRa, and Flk1 expression in Isl1-inducible ESCs at D4, 48 h in the presence or absence of Dox treatment. Percentages of cells in each quadrant are shown, and the percentage of CXCR4/PDGFRa/Flk1 TP cells are shown in parentheses. (C) FACS quantification of CXCR4/PDGFRa/Flk1 TP cells at 24 (D3) and 48 h (D4) in the presence or absence of Dox in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs. n = 4. (D) FACS quantification of cTNT expression at D8 in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs in the presence or absence of Dox from D2 to D4. n = 4. (E) Immunostaining of cTNT at D8 of differentiation in Dox-inducible Mesp1, Isl1, and Mesp1/Isl1 ESCs in the presence or absence of Dox from D2 to D4. Images shown are mosaïc acquisitions representative of at least four biologically independent experiments. Bars, 500 µm. (F) FACS quantification of CD31 expression at D7 in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs in the presence or absence of Dox from D2 to D4. n = 4. (G) Immunostaining for VE-Cadherin expression at D7 in Dox-inducible Mesp1, Isl1, and Mesp1/Isl1 ESCs in the presence or absence of Dox from D2 to D4. Images shown are representative of four biologically independent experiments. Bars, 100 µm. (H and I) FACS quantification of cTNT (H) and CD31 expression (I) in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs at D8 and D7 of differentiation, respectively, in the presence or absence of Dox from D2 to D4 or from D5 to D6. n = 4. Error bars indicate means ± SEM. TRE, tetracycline-responsive element.

Isl1 and Mesp1 cooperate in promoting cardiovascular differentiation in ESCs. (A) Schematic representation of Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs. (B) FACS analysis of CXCR4, PDGFRa, and Flk1 expression in Isl1-inducible ESCs at D4, 48 h in the presence or absence of Dox treatment. Percentages of cells in each quadrant are shown, and the percentage of CXCR4/PDGFRa/Flk1 TP cells are shown in parentheses. (C) FACS quantification of CXCR4/PDGFRa/Flk1 TP cells at 24 (D3) and 48 h (D4) in the presence or absence of Dox in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs. n = 4. (D) FACS quantification of cTNT expression at D8 in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs in the presence or absence of Dox from D2 to D4. n = 4. (E) Immunostaining of cTNT at D8 of differentiation in Dox-inducible Mesp1, Isl1, and Mesp1/Isl1 ESCs in the presence or absence of Dox from D2 to D4. Images shown are mosaïc acquisitions representative of at least four biologically independent experiments. Bars, 500 µm. (F) FACS quantification of CD31 expression at D7 in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs in the presence or absence of Dox from D2 to D4. n = 4. (G) Immunostaining for VE-Cadherin expression at D7 in Dox-inducible Mesp1, Isl1, and Mesp1/Isl1 ESCs in the presence or absence of Dox from D2 to D4. Images shown are representative of four biologically independent experiments. Bars, 100 µm. (H and I) FACS quantification of cTNT (H) and CD31 expression (I) in Mesp1, Isl1, and Mesp1/Isl1 Dox-inducible ESCs at D8 and D7 of differentiation, respectively, in the presence or absence of Dox from D2 to D4 or from D5 to D6. n = 4. Error bars indicate means ± SEM. TRE, tetracycline-responsive element.

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