Microarray profiling of pluripotent stem cell derived cardiomyocytes pretreated with DMSO, DHS (10µM final concentration) and co-incubation of DHS and curcumin (10µM final concentrations) for 3 consecutive days.
ABSTRACT: hCMs pretreated with DMSO : DMSO-1 and DMSO-2 hCMs pretreated with DHS: Sph1, Sph2 and Sph3 hCMs pretreated with DHS+ Curcumin: Sph+Cur1, Sph+Cur2 and Sph+Cur3
Project description:Investigation of whole genome gene expression level changes of Panc1 pretreated with Clofibrate, compared to Panc1 pretreated with DMSO. Compare differentially expressed genes by analyzing mRNA profilings between DMSO and Clofibrate pretreating Panc1 cell.
Project description:Pax5 controls the identity and development of B cells by repressing lineage-inappropriate genes and activating B-cell-specific genes. Here, we used genome-wide approaches to identify Pax5 target genes in pro-B and mature B cells. In these cell types, Pax5 bound to 40% of the cis- regulatory elements defined by mapping Dnase I hypersensitive (DHS) sites, transcription start sites and histone modifications. Although Pax5 bound to 8,000 target genes, it regulated only 4% of them in pro-B and mature B cells by inducing enhancers at activated genes and eliminating DHS sites at repressed genes. Pax5-regulated genes in pro-B cells account for 23% of all expression changes occurring between common lymphoid progenitors and committed pro-B cells, which identifies Pax5 as an important regulator of this developmental transition. Regulated Pax5 target genes minimally overlap in pro-B and mature B cells, which reflects massive expression changes between these cell types. Hence, Pax5 controls B cell identity and function by regulating distinct target genes in early and late B lymphopoiesis. 44 samples (16 RNA-seq, 15 ChIP-seq, 6 DHS-seq, 5 Bio-ChIP-seq, 2 CAGE-seq). All but four samples in in 2 biological replicates (8819, 8275, 8095, 8666). WT and experimental samples are provided.
Project description:Comparison of histone variant CENP-A in different cell lines vs. DHS profiles from same cell lines. Mock IPs included. Profiles of ectopic CENP-A IP after pre-clearing with CENP-B IPs, or profiles of DHS regions from mid-log culture SW480 colorectal cancer cell lines, HeLa cervical cancer cell lines, and EpiCo normal colon cell line were generated using deep sequencing after Mnase-based ChIP, in replicates, on the Illumina platform.
Project description:Phosphonate related fungicides such as neutralized phosphorous acid (NPA) are effective for the control of plant diseases caused by Oomycetes including Phytophthora parasitica. It has been proposed that phosphonate may induce plant resistance. However, the mechanism underlying phosphonate-induced resistance remains unclear. The purpose of this study is to identify genes that are differentially expressed in phosphonate-pretreated tomato plants in response to inoculation with Phytophthora parasitica. Overall design: We treated the tomato plants with NPA or water (ddH2O, as a control) for three days, and then inoculated the plants with zoospores of P. parasitica. For microarray analysis by the use of Affymetrix Tomato Genome Array, total RNAs were isolated from plant leaves at: (1) 3 days post treatment (dpt) with NPA or water, (2) 6 hours post inoculation (hpi) of the NPA- or water-pretreated plants, (3) 12 hpi of the NPA- or water-pretreated plants. To reduce the biological variability, each hybridization experiment was performed with RNA prepared from pools of five plants. The total set of experiments was repeated three times.
Project description:Distant enhancer elements are a major source of specificity in mammalian gene expression. Although enhancers that regulate broad developmental decisions and inducible gene expression have been studied extensively, little is known about regulatory elements that govern monogenic and monoallelic expression. Here, using high throughput epigenetic and genetic techniques we identified a plethora of distant enhancers that regulate monoallelic olfactory receptor (OR) gene expression. Potential OR enhancers have unique, cell type specific epigenetic marks that distinguish them from other neuronal enhancers and correlate with enhancer activity in vivo. Using sequence capture to enrich for these sequences we identified Dnase-protected footprints that reveal novel regulatory sequences and transcription factors required for OR gene activation. Our experiments provide insight to the regulation of OR expression, and describe novel principles and methodologies towards the understanding of transcriptional mechanisms that generate cellular diversity. In vivo examination of H3K79me3 enrichment and DNAse protected footprints on olfactory receptor enhancer sequences. We performed ChIP-seq on native chromatin isolated from the mouse olfactory epithelium using antibodies against H3K79me3. To sequence accessible regions of the genome we treated nuclei with limiting amounts of DNAse I to digest accessible chromatin and perform Dnase Hypersensitivity (DHS)-seq. In the olfactory epithelium, the H enhancer – the first described enhancer for olfactory receptors has a well-defined DNAse I hypersensitivity peak and is flanked by high levels of H3K79me3. We find other intergenic sequences nearby olfactory receptor genes that share the same chromatin signature, and test their function in vivo. TO uncover transcription factor footprints on olfactory receptor enhancers we performed sequence capture of the DHS-seq library to enrich for these sequences. We find multiple DNAse-protected sequences and perform motif analysis on transcription factor footprints to reveal factors involved in olfactory receptor gene regulation.
Project description:Transcriptional profiling of Curcumin treated S. mansoni adult worms (15uM of Curcumin in DMSO during 24h) vs. Non-treated S.mansoni adult worms (DMSO during 24h) Two conditions (treated vs. Non-treated) with four biological samples of treated worms (Curcumin 1, 2, 3 and 4) and four biological samples of non-treated worms (Control 1, 2, 3 and 4). For each sample (four treated and four non-treated) were performed two technical replicas.
Project description:The physiology of ethanologenic Escherichia coli grown anaerobically in alkaline-pretreated plant hydrolysates is not well studied. To gain insight into how E. coli responds to such hydrolysates, we studied an E. coli K-12 ethanologen fermenting a hydrolysate prepared from corn stover pre-treated by ammonia fiber expansion. Despite the high sugar content (~6% glucose, 3% xylose) and relatively low toxicity of this hydrolysate, E. coli ceased growth long before glucose was depleted. Nevertheless, the cells remained metabolically active and continued conversion of glucose to ethanol until all glucose was consumed. Gene expression profiling revealed complex and changing patterns of metabolic physiology and cellular stress responses throughout the different stages of growth. During the exponential and transition phases of growth, high cell maintenance and stress response costs were mitigated, in part, by free amino acids available in the hydrolysate media. However, after the majority of amino acids were depleted from the media cells entered stationary phase and ATP derived from glucose fermentation was consumed entirely by the demands of cell maintenance in the hydrolysate. Comparative gene expression profiling and metabolic modeling of the ethanologen suggested that the high energetic cost of mitigating osmotic, lignotoxin and ethanol stress collectively limits growth, sugar utilization rates and ethanol yields in alkaline-pretreated lignocellulosic hydrolysates. 38 samples in total. 24 samples were derived from biological replicate fermentations of alkaline-pretreated cornstover hydrolysate (12 datapoint time-series per fermentation). The remaining samples were obtained from fermentations conducted in defined media (Glucose Minimal Media (GMM, n=7), Synthetic Hydrolysate media (SynH, n=7)).
Project description:Adipogenesis is tightly controlled by a complex network of transcription factors acting at different stages of differentiation. Peroxisome proliferator-activated receptor gamma (PPAR gamma) and CCAAT/enhancer binding protein (C/EBP) family members are key regulators of this process. We have employed DNase I hypersensitive site analysis to investigate the genome-wide changes in chromatin structure that accompany the binding of adipogenic transcription factors. These analyses revealed a dramatic and dynamic modulation of the chromatin landscape during the first hours of adipocyte differentiation that coincides with cooperative binding of multiple early transcription factors (including glucocorticoid receptor, retinoid X receptor, Stat5a, C/EBPbeta and -delta) to transcription factor 'hotspots'. Our results demonstrate that C/EBPbeta marks a large number of these transcription factor 'hotspots' prior to induction of differentiation and chromatin remodeling and is required for their establishment. Furthermore, a subset of early remodeled C/EBP binding sites persists throughout differentiation and is later occupied by PPAR gamma , indicating that early C/EBP family members, in addition to their well established role in activation of PPAR gamma transcription, may act as pioneering factors for PPAR gamma binding. DNase I hypersensitive chromatin regions and transcription factor binding sites were identified at various time points of 3T3-L1 differentiation using DHS-seq and ChIP-seq, respectively.
Project description:The goal of this study was to identifiy cellular pathways modified by calcium storing in cells presenting a glycolytic metabolism and a poor mitochondrial biogenesis. Keywords: calcium stress response in RO82 W-1 cells Overall design: In duplicate, cells treated with ionomycin or BAPTA/AM during 96h compared to equivalent DMSO treated cells