Project description:Human aortic endothelial cells were stimulated by lysophosphatidylcholine (LPC) (10μM) with or without interleukin 35 (IL-35) (10ng/mL) or IL-10 (10ng/mL) for 18 hours. Total RNAs were extracted from samples, then mRNA and non-coding RNAs were enriched by removing rRNA from the total RNA. The library was sequenced by Illumina HiSeq4000 using PE100 strategy and the reads were mapped to the human hg19 reference genome.
Project description:To investigate the gene expression changes due to LPC-induced calcific nodule formation in porcine aortic valve interstitial cells (paVICs). We then examined differential gene expression due to treatment with dantrolene, which inhibits calcific nodule formation, with and without LPC after 24 hours of exposure.
Project description:To determine the molecular signaling pathways responsible for ETV4 regulation of cell growth, RNA sequencing was conducted on LPC-HRas cells with or without ETV4 knockdown
Project description:Rat OPCs were incubated in the presence of LPC 18:1 or LPC 18:0 and C13-His. Cultures controls included no treatment and a positive control for the differentiation of OPCs. LPC 18:1 was administered at 10 uM while C13-His was administered at 10 uM. Samples were process for mass spectrometry to identify the incorporation of C13-His in newly synthesized proteins.
Project description:Chemotherapeutic use of cisplatin is limited by its severe side effects. In this study, we demonstrated that cisplatin induces cell death in a proximal tubular cell line by suppressing glycolysis- and tricarboxylic acid (TCA)/mitochondria-related genes. HK-2 cells were cultured to confluence in 100mm dishes. Total RNA was extracted (QIAGEN, Valencia, CA, USA), and the concentration in the samples was measured using a Micro UV-Vis fluorescence spectrophotometer (Malcom, Tokyo, JAPAN). Sample of 10ɥg of Total RNA from HK-2 cells were labeled with biotin (3'IVT Labeling Kit, Affymetrix, USA) and hybridized (GeneAtlas Hybridization, Wash, and Stain Kit for 3' IVT Arrays, Affymetrix). The Microarray platform used was Affymetrix HG U219 Array Strip. The arrays were scanned in a GeneAtlas scanner controlled by GeneAtlas Software (Affymetrix, Sta. Clara, USA). Genes were later filtered according to expression fold change (Affymetrix Expression Console software, Affymetrix) Using Affymetrix GeneAtlas System, we determined the gene expression profiles of HK-2 cells treated with cisplatin. Two replicates were made for each timepoints (control, 6h and 24h).
Project description:Oligodendrocyte progenitor cells (OPC) were cultured in the presence of LPC 18:1-Cy5. Proteins interacting with LPC 18:1-Cy5 were UV crossed linked and cell lysates were separated via isoelectric focusing gel electrophoresis. Band was cut out from gel and processed for mass spectrometry. OPC lysates were also incubated with LPC 18:1-micelles and allowed to interact with OPC proteins. Samples were separated using sucrose gradient (Liposome floatation assay), top fraction was collected and processed for mass spectrometry.