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. Author manuscript; available in PMC: 2022 Jul 27.
Published in final edited form as: Nat Metab. 2022 Jan 27;4(1):123–140. doi: 10.1038/s42255-021-00514-4

Figure 5. Hemodynamic forces promote vMC coverage through oxPPP and ELN expression.

Figure 5

a, Effects of shear stress on the expression of G6PD and PGD genes exposed to 24hr of LSS (Laminar Shear Stress) in human endothelial cell lines (HAEC). KLF2 was used a positive control being a flow-induced gene. N=4 from three independent experiments. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test.

b, Effects of shear stress on the expression of different ECM target genes in HAEC after 24hr of LSS. n=4 from three independent experiments. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test.

c, qRT-PCR analysis of ELASTIN expression after LSS in G6PDkd EC. Scatter plots show the quantification of LSS/Static ratio in control and G6PDKD in HAEC. n=5 from four independent experiments. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test.

d, qRT-PCR analysis of pgd and g6pd expression in ECs sorted from Tg(kdrl:eGFP)s843 treated with BDM 4mM or control at 72 hpf. n=4 from two independent experiments. Fifty GFP-positive embryos from Tg(kdrl:eGFP)s843 were used in each sample. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test.

e, qRT-PCR analysis of elna and elnb expression in ECs sorted from Tg(kdrl:eGFP) embryos treated with BDM or injected with control or gata1 MO at 72hpf. Ctrl and gata1a MO (n=3), BDM (n=4) from two independent experiments. Fifity GFP-positive embryos from Tg(kdrl:eGFP)s843 were used in each sample. Data are shown as mean ± SEM. Statistics were done using one-way ANOVA followed by Dunnet’s multiple comparison test.

f, Confocal images of partial z-projection of the trunk region (somite 8-14) of a Tg(kdrl:eGFP)s843 ;Tg(acta2:mCherry)uto5 embryo at 4dpf after elna and elnb antisense morpholino (MO) injection. Scale bar, 100 μm.

g, Scatter plots show the defect in vMC coverage in elna/elnb morphants at 4dpf. Ctrl (n=11), elna MO (n=7), elnb MO (n=10), elna/elnb MO (n=9) embryos from 2 independent experiments. Data are shown as mean ± SEM. Statistics were done using one-way ANOVA followed by Tukey’s multiple comparison test.