ABSTRACT: To know effects of long noncoding RNA (lncRNA) SCIRT in hypoxia/reoxygenation (H/R). We compared expression profiles of human umbilical vein endothelial cells (HUVECs) under normal condition, H/R condition without SCIRT knockdown, and H/R condition with SCIRT knockdown.
Project description:To have a global view of transcriptional change of hypoxia/reoxygenation (H/R) condition compared with normal condition, we collected human umbilical vein endothelial cells (HUVECs) from both conditions. The expression profiles of HUVECs were detected by microarray, and two conditions were compared to detect significantly changed lncRNAs and mRNAs.
Project description:Human umbilical vein endothelial cells (HUVEC) were cultivated on 1%-gelatin-coated 75-cm2 culture flasks in Medium 200 supplemented with 2% fetal calf serum, penicillin (100 units/ml), streptomycin (100 units/ml) and Funizone (0.25 μg/ml) supplied by Cascade Biologics (Portland, OR). Prior to an experiment, HUVECs were subcultivated with Trypsin/EDTA onto 1% gelatin coated glass 30-mm glass coverslips in 60 mm glass Petri dishes. The hypoxic treatment was performed in airtight chambers from Mitsubishi Gas Chemical Co. Inc (Remel Scientific, Baton Rouge, LA) Preconditioning consisted of 1-hour hypoxia followed by various periods of reoxygenation (0, 3, 5, 12 and 24 hrs) for gene expression analysis. Total RNA was extracted from cultured HUVECs with TRITM reagent according to the manufacture’s instructions (Molecular Research Center, Cincinnati, OH). Cy5 and Cy3 labeled cDNA probes were generated from total RNA by reverse transcription in the presence of aminoallyl-dUTP according to the method described by Xiang, et al,15. Human universal reference RNA (Stratagene, La Jolla, CA) was Cy3-labeled and used as reference RNA and RNA isolated from HUVEC at 0, 3, 5, 12 and 24 hour following reoxygenation were Cy5-labeled. The high-resolution scans of hybridized microarrays were acquired with a GenePix scanner 4000B (Axon Instruments, Inc., Union City, CA) and tabulated with GenePix pro 5.1 software. Keywords: time-course
Project description:Human umbilical vein endothelial cells (HUVECs) were incubated for 48 h under normoxic or hypoxic (1% oxygen) conditions. Changes in transcript and exon levels were analyzed. 6 samples; 2 conditions: normoxia vs. hypoxia (1% oxygen); 3 replicates per condition
Project description:Human umbilical vein endothelial cells (HUVECs) were incubated for 48 h under normoxic or hypoxic (1% oxygen) conditions. Changes in transcript and exon levels were analyzed.
Project description:Biochemical characterization of the hypoxia-induced long non coding RNA NTRAS. NTRAS was found to regulate cell cycle progression, in vitro sprouting angiogenesis, and the paracellular permeability in human umbilical vein endothelial cells (HUVECs). Using desthiobiotinylated 2’O-Me-RNA probes, we purified endogenous NTRAS-protein-complexes and identified NTRAS interacting proteins by mass spectrometry.
Project description:Human umbilical vein endothelial cells (HUVECs) were insulted with cobalt chloride to induce cell apoptosis and alternative splicing in the mimic-hypoxia environment. We use Affymetrix exon array to reveal differential expression from transcript-level and exon-level in genome-wide.
Project description:To understand how plants respond to anoxia-reoxygenation, we have employed a global microarray expression profiling as a basic platform to characterize genes with the potential to mediate the recovery responses in Arabidopsis. 7-day-old seedlings were trated with 4hr and 8hr anoxia, and then reoxygenated in air for 0, 0.5, 1, 3, 6 hrs. Genes changed in both condition were designated as reoxygenation-regulated genes. They included the genes in ROS detoxification, dehydration, metabolic process, and many other responses. Interestingly, ethylene was also involved in this recovery process. We further adopted ein2-5 and ein3eil1 microarray to investigate ethylene signaling. Our results showed ethylene partially regulate reoxygenation regulated genes and is reruired for plant survival in reoxygenation. Genes expression during anoxia-reoxygenation in Arabidopsis seedlings was measured at 0, 0.5, 1, 3, 6 hours after anoxia treatment, and normal condition before anoxia. Three independent experiments were performed at each time (Nor, 0, 0.5, 1, 3, 6) under 4hr anoxia-reoxygenation condition by using Col-0, ein2-5, and ein3eil1. Four independent experiments were performed at each time (Nor, 0, 0.5, 1, 3, 6) under 8hr anoxia-reoxygenation condition by using Col-0.
Project description:To investigate machanism of miR-210-3p regulating angiogenic ability of human umbilical vein endothelial cells (HUVECs) in hypoxic conditions, we transfected miR-210-3p mimic to overexpress miR-210-3p in human umbilical vein endothelial cells. We than performed RNA sequencing of miR-210-3p mimic-transfected and control HUVECs under hypoxic conditions to evaluate the transcriptional changes in the miR-210-3p-overexpressing HUVECs.
Project description:We quantified differential microRNA (miRNA) expression in Human umbilical vein endothelial cells (HUVECs)response to Angiogenin (ANG) treatment.These data were used to determine which miRNAs are altered on ANG in Human umbilical vein endothelial cells.