RNAseq data from paper: The loss of DHX15 impairs endothelial energy metabolism, lymphatic drainage and tumor metastasis in mice

RNA from non-silenced and DHX15-silenced LECs was extracted (n=4 for experimental condition). The transcriptome analysis was carried out using RNAseq (HiSeq, Illumina). Total RNA from <i>Mus musculus</i> was quantified by Qubit® RNA BR Assay kit (Thermo Fisher Scientific) and the RNA integrity was estimated by using RNA 6000 Nano Bioanalyzer 2100 Assay (Agilent). The RNASeq libraries were prepared with KAPA Stranded mRNA-Seq Illumina® Platforms Kit (Roche-Kapa Biosystems) following the manufacturer´s recommendations. 500ng of total RNA was used as the input material, the poly-A fraction was enriched with oligo-dT magnetic beads and the mRNA was fragmented. The strand specificity was achieved during the second strand synthesis performed in the presence of dUTP instead of dTTP. The blunt-ended double stranded cDNA was 3´-adenylated and Illumina indexed adapters (Illumina) were ligated. The ligation product was enriched with 15 PCR cycles and the final library was validated on an Agilent 2100 Bioanalyzer with the DNA 7500 assay. The libraries were sequenced on HiSeq 4000 (Illumina, Inc) in paired-end mode with a read length of 2x76bp using HiSeq 4000 SBS kit in a fraction of a HiSeq 4000 PE Cluster kit sequencing flow cell lane, following the manufacturer’s protocol. Image analysis, base calling and quality scoring of the run were processed using the manufacturer’s software Real Time Analysis (RTA 2.7.6) and followed by generation of FASTQ sequence files by CASAVA. RNA-seq reads were mapped against the mouse reference genome (GRCm38) using STAR version 2.5.2a⁶⁸ with ENCODE parameters for long RNA. Genes were quantified with RSEM version 1.2.28⁶⁹ using the gencode M12 version. <br>