Transcription analysis of photocoagulated human retinal pigment epithelial cells in culture
ABSTRACT: ARPE-19 cells were either laser treated or not, and analyzed 2, 6 and 24 hours after laser treatment. In addition, the cells were treated with high or low glucose (T2D complications analysis). The difference in glucose levels did not make much difference in regards to gene expression. 52 ARPE-19 samples including controls and laser-treated samples. Separate analysis for high and low glucose levels.
Project description:NF1-C2 suppresses tumorigenesis and epithelial-to-mesenchymal transition by repressing FoxF1. We used microarray to identify direct targets for NF1-C2. The human breast tumor cell line MDA-MB-436 were stably transfected with an expression plasmid including NF1-C2. RNA from these cells and wildtype cells were extracted and hybridized on Affymetrix microarrays (platform GPL570). Three replicates each.
Project description:This experiment aimed to measure gene expression in glucose-responsive beta-cells. RNA was extracted using RNeasy Mini kit. Quantity and quality of extracted RNA were assessed by the NanoDrop spectrophotometer ND-1000 and Experion RNA StdSens Analysis Kit, respectively. Background correction, normalization, and probe summarization were performed by the Robust Multichip Average method. More details can be found in J Biol Chem (doi: 10.1074/jbc.M112.422527).
Project description:We used microarrays to evaluate the effect of SRPIN803 on gene expression in ARPE-19 cells. ARPE-19 cells were treated with SRPIN803 (10 uM) or the negative control (0.1% DMSO) for 4 hours for Total RNA isolation and hybridization on Microarray.
Project description:To characterize the potential molecular pathway(s) affected by iron treatment and identify the one(s) responsible for C3 induction, we performed a whole genome microarray on untreated ARPE-19 cells and cells treated with 250 μM FAC for 48h/2d. Gene expression was compared between untreated and FAC-treated ARPE-19 cells, with three biological replicates in each.
Project description:Background. Differential gene expression in adipose tissue during diet-induced weight loss followed by a weight stability period is not well characterized. Markers of these processes may provide a deeper understanding of the underlying mechanisms. Objective. To identify differentially expressed genes in human adipose tissue during weight loss and weight maintenance after weight loss. Design. RNA from subcutaneous abdominal adipose tissue from nine obese subjects was obtained and analyzed at baseline, after weight reduction on a low calorie diet (LCD), and after a period of group therapy in order to maintain weight stability. Results. Subjects lost 18.8 + 5.4% of their body weight during the LCD and maintained this weight during group therapy. Insulin sensitivity (HOMA) improved after weight loss with no further improvement during weight maintenance. Cyclin-dependent kinase inhibitor 2B (CDKN2B) and JAZF zinc finger 1 (JAZF1), associated with type 2 diabetes, were downregulated. We could also confirm the downregulation of candidates for obesity and related traits, such as tenomodulin (TNMD) and matrix metallopeptidase 9 (MMP9), with weight loss. The expression of other candidates, such as cell death-inducing DFFA-like effector A (CIDEA) and stearoyl-CoA desaturase (SCD) were upregulated during weight loss but returned to baseline levels during weight maintenance. Conclusion. Genes in the adipose tissue are differentially expressed during weight loss and weight maintenance after weight loss. Genes that show sustained regulation may be of potential interest as markers of the beneficial effects of weight loss whereas others seem to be primarily involved in the process of weight loss itself. Nine participants were prescribed a low calorie diet (LCD) containing 1200 kcal/day for approximately three months (101 ± 26 days). Following the weight reduction phase the participants attended a six month follow-up period (167 ± 37 days). By protocol design, subjects were eligible to enter the study if they had lost at least 10% of their initial body weight during the LCD-period and maintained this weight (+5%) after group therapy. Subcutaneous adipose tissue samples were obtained at three time-points: (i) at baseline, (ii) after weight reduction when subjects were no longer losing weight, and (iii) after the group therapy weight maintenance phase.
Project description:To investigate the effects of quality of fat in a high fat diet (HFD) over time on hepatic lipid storage and transcriptome in mice. In this dataset, we include the expression data obtained from dissected mouse liver after being fed with Control, HFD-EPA/DHA and HFD-corn oil diet for 8 and 12 weeks. In total, 24 samples were analyzed. Based on hierarchical clustering and scatter plots, one microarray from the HFD-corn oil group at 8 weeks was removed from subsequent analyses. The Piano package was used for Gene Set Enrichment Analysis (GSEA) to identify the total number of genes regulated and the direction of regulation. Gene Ontology (GO) terms were annotated to each probe-set after performing GSEA. The reporter algorithm was used to analyse the functional enrichment level of individual GO term. Heatmaps were generated using the log10p-values to visualize enriched GO biological processes (BPs) terms.
Project description:We have performed a proteomics analysis of a human retinal pigment epithelial cell line (ARPE-19), which represents a widely used model for in vitro studies of cellular and molecular mechanisms related to human RPE cells. The project was jointly supervised by <b>Francesco Giorgianni</b> and <b>Sarka Beranova-Giorgianni</b>.
Project description:LDL or Ox-LDL 200ug/ml, which showed no loss of viability after a 48 hour exposure, induced a physiological and pathological transcriptional response, respectively. LDL induced a downregulation of genes associated with cholesterol biosynthesis while ox-LDL induced transcriptional alterations in genes related to inflammation, matrix expansion, lipid metabolism and processing, and apoptosis. Pentraxin-3 was secreted into the culture medium after RPE cells were stimulated with ox-LDL, and immunohistochemically evident in Bruchs membrane of human macular samples with age-related macular degeneration. ARPE-19 cells exposed to 200ug/ml ox-LDL had a 38% apoptosis rate compared to less than 1% when exposed to LDL or untreated controls (p<0.0001). While LDL induced a physiologic response by RPE cells, a pathological phenotypic response was seen after treatment with oxidatively modified LDL. The transcriptional, biochemical, and functional data provide initial support of a role for the hypothesis that modified LDLs are one trigger for initiating events that contribute to the development of age-related macular degeneration. Keywords: treatment with non-treatment control Human ARPE-19 cells were exposed to LDL or oxidatively modified LDL (ox-LDL) for 48 hours for RNA extraction and hybridization on Affymetrix microarrays. We sought to determine whether retina, pigment epithelial cells develop a pathologic phenotype after exposure to low density lipoproteins (LDL) that are oxidatively modified.We have made two comparsions: LDL treatment versus non-treatment; ox-LDL treatment versus non-treatment.