Fig. 1.

LINC00607 is an EC-enriched lncRNA upregulated during hypoxia and EndMT. A FANTOM5 CAGE-ENCODE expression of the 9 highest endothelial expressed lncRNAs across different cell lines. Each cell type-specific signal was divided through the mean signal observed in all cell types. B FANTOM5 CAGE expression of the 9 highest expressed endothelial lncRNAs across different endothelial tissues. Calculation was performed as in A. C RNA-FISH of LINC00607 in HUVEC. LINC00607 is labelled with a 5’TYE-665 probe, DAPI is used to stain the nuclei. Scale bar indicates 20 µm. D, E RNA-FISH of LINC00607 in cultured endothelial cells retrieved from human plaque-free (healthy, free) and plaque-containing (atherosclerosis, cont.) arteries. Scale bar indicates 20 µm. Fluorescence intensity per cell for TYE-665 (LINC00607) is shown in E. n = 26–30, unpaired t test. F RT-qPCR of LINC00607 in human arteriovenous malformations (AVM) treated with and without the β-blocker propranolol for 72 h. n = 5, Mann–Whitney U test. G RT-qPCR of the LINC00607 homologue in vessels originating from Macaca fascicularis treated either with a normal diet (CTL), a high fat diet (Ath) or with a high-fat diet and a subsequent recovery phase (Reg). n = 3. One-way ANOVA with Bonferroni post hoc test. H RNA in situ hybridization of carotid arteries from Macaca fascicularis (Mf) with RNAscope. Arrows point to dots indicating LINC00607. Representative images are shown. Scale bar indicates 10 µm. I Promoter analysis of LINC00607. A region starting from the LINC00607 transcriptional start site (TSS) and 1000 base pairs (bp) upstream was analyzed with MoLoTool and plotted according to p value. J Relative LINC00607 expression in HUVEC treated with normoxia (NOX) or hypoxia (HOX), n = 7. Mann–Whitney t test. K LINC00607 gene read counts in HUVEC cultured under normoxic and hypoxic conditions, n = 3. Unpaired t test. L Volcano plot of log2 fold changes of lncRNAs expressed in hypoxia versus normoxia. M Relative expression of LINC00607 in HUVEC after stimulation with DMOG. DMSO served as control (CTL), n = 8, Mann–Whitney U Test. N Relative expression of LINC00607 in HUVEC after stimulation with acriflavine (ACF), n = 7, Mann–Whitney U Test. O, P Volcano plot of log2 fold changes of lncRNAs in HUVEC treated with acriflavine (ACF) cultured under normoxia (O) or hypoxia (P). Q Relative expression of LINC00607 in HUVEC under basal (CTL) or Endothelial-to-mesenchymal transition (EndMT) conditions. n = 3, Unpaired t test. R, S Volcano plot of log2 fold changes of lncRNAs after EndMT versus unstimulated control in HUVEC (R) or pulmonary arterial endothelial cells (PAEC) (S). Error bars are defined as mean ± SD. *p < 0.05