Extended Data Fig. 8: LSS-dependent expression of G6PD, PGD and ELN in EC.

a, Effects of shear stress on the expression of G6PD and PGD genes exposed to 24h of LSS in human endothelial cell lines (HUVEC). KLF2 was used a positive control being a flow-induced gene. KFL2 (n=3), G6PD and PGD (n=6) from three independent experiments. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test. b, Shear stress increases G6PD and PGD expression in ECs. Laminar shear stress (LSS) induces oxPPP in pulsatile shear flow (PS) and in oscillatory shear flow (OS). Longitudinal transcriptional response of G6PD and PGD genes (10 time points, 42 RNAseq samples). Gene counts were obtained from fastq data from SRP117215 dataset of SRA ⁴⁰ quantified using rsem and normalized with EDASeq. Data are shown as mean ± SEM. c, Effects of shear stress on the expression of different ECM target genes in human endothelial cell lines (HUVEC) after 24h of LSS. n=4 from three independent experiments. Data are shown as mean ± SEM. Statistics were done using unpaired Student’s t-test. d, (Top) Generation of CRISPR vectors for EC-specific gene targeting. Schematic representation, adapted from Ensembl, illustrating the exons in elna gene and the gRNA target sequence used to generate endothelial-specific KO for elna. The CRISPR targeted sequence is underlined. The red letters indicate the PAM region for each gene. (Bottom) Heteroduplex mobility assay of the PCR-amplified elna exon 5 fragment targeted by the CRISPR/Cas9 sgRNA. 1,2, and 3 represent three sgRNA-injected embryos; wt1 and wt2 are control embryos. e, Scatter plots showing the quantification of vMC coverage of Tg(kdrl:eGFP)^s843 Tg(acta2:mCherry)^uto5 embryos at 4dpf in g6pd^wt and g6pd^sa24272 with and without elna mRNA injected at 100ng/μl. g6pd^wt (n=6), g6pd^sa24272 (n=9), g6pd^wt + elna 100ng/μl (n=6), g6pd^sa24272 + elna 100ng/μl (n=10) embryos from two independent experiments. Data are shown as mean ± SEM. Statistics were done using one-way ANOVA followed by Tukey’s multiple comparison test.

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