Investigation of Gene Expression Changes in Human Umbilical Vein Endothelial cells (HUVECs) following SLC25A13 Knockdown via shRNA Lentiviral Transduction

In this study, we discover an essential role for the mitochondrial glutamate-aspartate antiporter SLC25A13 in biomass synthesis in endothelial cells (ECs). To delineate the global changes of SLC25A13 silencing at a molecular level, we performed high-throughput bulk RNA-sequencing and analyzed the differences between the gene expression profiles of CTRL and SLC25A13-KD ECs. HUVECs were transduced at a multiplicity of infection (MOI) of 30. Transductions were performed on day 0 and after 24 hours the cells were refed with fresh medium. RNA of scrambled control and SLC25A13-KD ECs were extracted 6 days after seeding. Samples were indexed to allow for multiplexing. Libraries were sequenced on an Illumina NovaSeq6000 platform. Single-end reads of 100 bp length were produced with a minimum of 20 M reads per sample. The reads were aligned using a standard pipeline for splice-aware alignment. The reads per gene were then quantified. The study accounted for 12 samples (6 for CTRL and 6 for SLC25A13-KD ECs).