|
AIBP augments LEC fate commitment. a Representative confocal images of Prox1+ cells in the fli1a:egfp and apoa1bp2−/−; fli1a:egfp at 4 dpf following immunostaining using GFP and Prox1 antibodies. Arrows show the Prox1+ LECs in TD. Blood vessels are labeled in green, and Prox1 staining is shown in red. Data are pooled from 3 independent experiments. b Quantitative data of Prox1+ LEC in TD of 7 somites. Mean ± SE, n = 23 (fli1a:egfp) and n = 28 (apoa1bp2−/−; fli1a:egfp) embryos; unpaired two-sided t-test with Welch’s correction. c Scheme illustration of mESC to LEC differentiation. The embryoid bodies (EBs) of mESCs were prepared and cultured in the EC differentiation medium containing BMP4 and bFGF for 3 days. Recombinant VEGFA and VEGFC in combination or AIBP were supplemented at day 3 and remained in culture for additional 4 days. d qPCR analysis of the expression of LEC-associated Lyve1 and Prox1 and endothelial cell marker Pecam1 at the indicated time points of mESC to LEC differentiation. Mean ± SD, n = 3 independent repeats; two-way ANOVA with Tukey’s post-hoc test. The P-values comparing control and AIBP-treated samples are shown. e qPCR analysis of the expression of Vegfc and Adamts3. Mean ± SD, n = 3-4; two-way ANOVA with Tukey’s post-hoc test. f Effect of VEGFR3 and NOTCH inhibition on mESC to LEC differentiation. The murine ESCs were subjected to LEC differentiation as described in panel c in the presence or absence VEGFR3 inhibitor SAR (SAR13165) or NOTCH inhibitor DAPT from day 3 to day 7. qPCR analysis of Prox1, Lyve1, and Pecam1 expression was performed. Mean ± SD, n = 3 independent repeats; two-way ANOVA with Tukey’s post-hoc test. ****p < 0.0001. Scale: 50 µm. Source data are provided as a Source Data file.
|
