RNA sequencing of wildtype-KRAS HKE3 (wtHKE3) and mutant-KRAS HKE3 (mtHKE3) colorectal cancer cell lines before and after stimulation with TGF-alpha
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE110649 | GEO | 2019/09/18
REPOSITORIES: GEO
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Project description:RNA sequencing was performed on HCT116 and HKE3 colorectal cancer cell lines before stimulation and at 15, 30, 60, 90 and 120 mins post-stimulation with transforming growth factor alpha (TGF-α) (0.01 µg/mL, Abcam). Three replicates were sequenced at each time point.
2019-09-18 | GSE105094 | GEO
Project description:This study involves the analysis of surgically resected samples from patients undergoing breast surgery. The samples published here have been used as part of the publication by [1], and all experimental details can be found within. Desorption electrospray ionisation mass spectrometry imaging (DESI MSI) was used to provide spatially resolved metabolic profiles across the various tissue samples, which in combination with histological assessment allows characterisation of changes across distinct tissue types. In being able to characterise the metabolic differences between tissue types, it becomes possible to predict future unknown samples using a data-driven histological approach to enhance current disease prognoses. </p> Ref:</br> [1] Guenther S, Muirhead LJ, Speller AV, Golf O, Strittmatter N, Ramakrishnan R, Goldin RD, Jones E, Veselkov K, Nicholson J, Darzi A, Takats Z. Spatially resolved metabolic phenotyping of breast cancer by desorption electrospray ionization mass spectrometry. Cancer Res. 2015 May 1;75(9):1828-37. doi:10.1158/0008-5472.CAN-14-2258. PMID:25691458
2016-04-28 | MTBLS273 | MetaboLights
Project description:PolyA+ RNA-Seq was performed on wildtype, ATRX and DAXX knockout cells derived by CRISPR targeting (Sadic et. al, EMBO Rep, 16(7), p836-850, 2015, DOI: 10.15252/embr.201439937). PolyA+ RNA-Seq was performed on wildtype, ATRX and DAXX knockout cells derived by CRISPR targeting (Sadic et. al, EMBO Rep, 16(7), p836-850, 2015, DOI: 10.15252/embr.201439937). Cells (isogenic, male)
2015-10-26 | E-GEOD-73881 | biostudies-arrayexpress
Project description:Comparative Dynamic Transcriptome Analysis (cDTA) enables global analysis of newly synthesized RNA as described in Sun et al. Genome Res. 2012 (DOI:10.1101/gr.130161.111) and reveals defects in transcription with much higher sensitivity than conventional steady-state methods. cDTA was carried out as described in Sun et al. Genome Res. 2012 (DOI:10.1101/gr.130161.111), using the S. cerevisiae heterozygous Med17/med17delta strain (Euroscarf) transfected with plasmids pRS315-SRB4 or pRS315-srb4-ts as described in Larivi̬re et al. Nature. 2012 (DOI:10.1038/nature11670), and Y40343-wildtype (Euroscarf) or Med18-FRB-KanMX6 (Euroscarf) strains. Heatshock of SRB4 and srb4-ts strains was applied for 18 or 60 minutes at 37C prior to RNA labeling as described in Sun et al. Genome Res. 2012 (DOI:10.1101/gr.130161.111). To deplete the Med18 subunit from the nucleus, anchor-away experiments were performed by rapamycin treatment (1 ug/ml in 200 mL YPD) for 18 or 60 minutes at 30C prior to RNA labeling as described in Sun et al. Mol. Cell. 2013 (DOI:10.1016/j.molcel.2013.09.010). Data analysis was as described in Sun et al. Genome Res. 2012 (DOI:10.1101/gr.130161.111).
2015-02-04 | E-MTAB-2942 | biostudies-arrayexpress
Project description:Loss of ultrafiltration capacity in peritoneal dialysis (PD) patients is often associated with peritoneal vascular changes, manifest as diabetes-like vasculopathy and angiogenesis. The endothelial monolayer lining the vessel lumen controls vessel barrier function and thereby influences peritoneal substrate transport and ultrafiltration. In this study, we investigated the influence of exposure of human umbilical vein endothelial cells (HUVEC) to different PD fluids. HUVECs were exposed to experimental solutions for up to 24 h. All test fluids were sterile-filtered before usage. Each experiment consisted of three independent samples in biological replicates on separate culture plates. For PD fluid incubation, cells were first exposed to pure PD fluids (Dianeal PD4 3.86% glucose, Physioneal 3.86% glucose, Extraneal, all Baxter, Castlebar, Ireland), or to a lab-made GDP-free PD fluid or to normal medium without growth factors as control. Cells were subsequently exposed for 24 h to the above solutions diluted 1:1 with culture medium and brought to 2% FCS. Cells were then washed three times (250 mM sucrose, 10 mM Tris/HCl, pH 7) and lysed in 125 µL per well of lysis buffer (30 mM Tris, pH 8.5, 7 M urea, 2 M thiourea, 4% CHAPS, 1 mM EDTA, one tablet of Complete Protease Inhibitor (Roche, Basel; Switzerland) per 100 ml, and one tablet of PhosStop Protease Inhibitor (Roche) per 100 mL). Total protein concentration was determined with the Pierce 660 Kit (Thermo) per manufacturer’s manual. Lysates were stored at -80°C until further processing. We investigated the molecular mechanisms of endothelial cell pathology during in vitro PD fluid exposure by proteomic and bioinformatic analysis of the endothelial cell response to the PD fluid induced stress, integrating current understanding of PD-related cellular injury and stress responses (Bender et al., Nephrol Dial Transplant, 2011, doi:10.1093/ndt/gfq484; Kratochwill et al., BioMed research international, 2015, doi:10.1155/2015/628158; Kratochwill et al., J Proteome Res, 2009, doi:10.1021/pr800916s; Lechner et al., J Proteome Res, 2010, doi:10.1021/pr9011574). We then evaluated the effect of AlaGln addition to conventional PD fluid on these parameters in endothelial cells. This strategy allowed characterization of endothelial cell injury and stress responses during exposure to different PD fluid.
2022-08-11 | PXD034985 | Pride
Project description:The aim of the experiment was to identify early gibberellin (GA) responsive genes in the roots of an Arabidopsis GA deficient mutant.The GA deficient mutant used in this study is a transgenic line overexpressing the PcGA2ox1 gene. This mutant has an identical phenotype to ga1-3, but it does not require exogenous GA treatment for germination.Seeds were germinated on 1xMS + 1% (w/v) sucrose plates containing 0.7% gelrite, and grown under continous light. The plates were orientated vertically.After six days growth the plates were treated with or without 5uM GA4 for 0, 30, 60 and 180 minutes.Approximately 400 hypocotyls were harvested per timepoint using a razorblade and after excision the hypocotyls were frozen in liquid nitrogen giving a total of 7 experimental samples (1: untreated, 2: 30 mins GA4, 3: 30 mins untreated, 4: 60 mins GA4, 5: 60 mins untreated, 6: 180 mins GA4, 7: 180 mins untreated. Three biological replicates were performed giving a total of 21 root samples. Total RNA was isolated from the roots using the QIAGEN RNeasy method. Keywords: Expression profiling by array
2009-11-18 | GSE18985 | GEO
Project description:We previously established a method to generate myeloid lineage cells with proliferation capacity from induced pluripotent stem cells (iPSCs) (Zhang R. Cancer Immunol Res. 3: 668, 2015). In this study, we generated the IFN-α-producing cells by genetic engineering of mouse iPSC-derived myeloid cells (iPSC-pMCs). Gene expression profiling of IFN-α-producing iPSC-pMCs (IFN-α-iPSC-pMCs) revealed a distinct interferon-stimulated gene (ISG) signature: of 613 differentially expressed genes (versus iPSC-pMC), 345 genes were ISGs of which > 70 % were type I IFN-related. Gene ontology analysis showed that the up-regulated gene signature was enriched for transcripts associate with immune responses, cellular responses to type I IFN, and defense responses to virus.
2019-02-13 | GSE126277 | GEO
Project description:Loss of ultrafiltration capacity in peritoneal dialysis (PD) patients is often associated with peritoneal vascular changes, manifest as diabetes-like vasculopathy and angiogenesis. The endothelial monolayer lining the vessel lumen controls vessel barrier function and thereby influences peritoneal substrate transport and ultrafiltration. In this study, we first characterized the response to conventional PD fluids of endothelial cells isolated from peritoneal biopsies of children undergoing PD. 9 omental biopsies from the pediatric biopsy biobank (mean age 5.7 years) were included in this study. PD patients were treated with conventional PD fluid (mean PD vintage 12.5 months). Age-matched controls (n=5) with normal renal function undergoing elective surgery were also included. In patients on PD, the biopsy sampling site was at least 5 cm away from the PD catheter entry site. Written informed consent was obtained from parents, and from patients as appropriate. The study was performed according to the Declaration of Helsinki and registered at www.clinicaltrials.gov (NCT01893710). The study was part of the International Pediatric Dialysis Network (IPDN; www.pedpd.org). Patients with a BMI of >35 kg/m2 and with chronic inflammatory diseases were excluded. Specimens obtained during surgery were instantaneously frozen with liquid nitrogen and stored at -80°C. Arterioles macroscopically located within fat tissue and microscopically at least 1mm distant from the peritoneal surface were micro-dissected. Elastica van Gieson stainings of the arteriolar elastic lamina were used as templates for microdissection of arterioles from cresol violet–stained neighboring sections. 100 µm thick tissue slices were cut with a cryotome and 30 deep-frozen arteriolar rings per patient were micro-dissected using a stereo microscope and a microneedle. Arteriolar rings were lysed in 100 µl lysis buffer (30 mM Tris, pH 8.5, 7M urea, 2M thiourea, 4% 3-[(3-Cholamidopropyl) dimethylammonio]-1-propanesulfonate (CHAPS), 1 mM EDTA, 1 tablet of Complete Protease Inhibitor (Roche, Basel, Switzerland) and 1 tablet of phosphatase inhibitor (PhosSTOP, Roche) per 100 ml). The resulting lysates were stored at -80°C until further processing. Total protein concentration was determined with the 2D-Quant kit (GE). Quality control and verification of protein concentrations was performed by SDS-PAGE. We investigated the molecular mechanisms of endothelial cell pathology during in vivo and in vitro PD fluid exposure by proteomic and bioinformatic analysis of the endothelial cell response to hyperglycemic stress, integrating current understanding of PD-related cellular injury and stress responses (Bender et al., Nephrol Dial Transplant, 2011, doi:10.1093/ndt/gfq484; Kratochwill et al., BioMed research international, 2015, doi:10.1155/2015/628158; Kratochwill et al., J Proteome Res, 2009, doi:10.1021/pr800916s; Lechner et al., J Proteome Res, 2010, doi:10.1021/pr9011574).
2021-09-09 | PXD022183 | Pride
Project description:PolyA+ RNA-Seq was performed on wildtype, ATRX and DAXX knockout cells derived by CRISPR targeting (Sadic et. al, EMBO Rep, 16(7), p836-850, 2015, DOI: 10.15252/embr.201439937).
2015-10-26 | GSE73881 | GEO
Project description:Loss of ultrafiltration capacity in peritoneal dialysis (PD) patients is often associated with peritoneal vascular changes, manifest as diabetes-like vasculopathy and angiogenesis. The endothelial monolayer lining the vessel lumen controls vessel barrier function and thereby influences peritoneal substrate transport and ultrafiltration. The dipeptide alanyl-glutamine (AlaGln) has recently shown cytoprotective effects when added to PD fluid, including preservation of mesothelial cells in vitro (Kratochwill et al., Nephrol Dial Transplant, 2012, doi:10.1093/ndt/gfr459) and attenuation of the exuberant angiogenesis seen in long-term rodent models of PD (Ferrantelli et al., Kidney international, 2016, doi:10.1016/j.kint.2015.12.005). These effects have been reflected in early phase clinical trials as restoration of effluent cell stress responses and improved biomarkers of peritoneal health (Kratochwill et al., PLoS One, 2016, doi:10.1371/journal.pone.0165045; Vychytil et al., Kidney international, 2018, doi:10.1016/j.kint.2018.08.031). In a randomized clinical phase II trial, 8 weeks treatment with AlaGln in PD fluid decreased peritoneal protein loss. This clinically important effect might be explained by preserved peritoneal membrane and vessel integrity, but the role of AlaGln in preservation of peritoneal endothelial cell function remains unclear. In this study, we investigated to what extent in vivo observed biological processes and pathways are replicated by in vitro exposure of human umbilical vein endothelial cells (HUVEC) to conventional PD fluid. HUVECs were exposed to experimental solutions for up to 24 h. All test fluids were sterile-filtered before usage. Each experiment consisted of three independent samples in biological replicates on separate culture plates. For PD fluid incubation, cells were first exposed for 1 h to pure glucose-based PD fluid (Dianeal PD4 3.86% glucose, Baxter, Castlebar, Ireland), or to the same PD fluid supplemented with AlaGln dipeptide (8 mM, Dipeptiven; Fresenius Kabi, Bad Homburg, Germany), or to normal medium without growth factors as control. Cells were subsequently exposed for 24 h to the above solutions diluted 1:1 with culture medium and brought to 2% FCS. Cells were then washed three times (250 mM sucrose, 10 mM Tris/HCl, pH 7) and lysed in 125 µL per well of lysis buffer (30 mM Tris, pH 8.5, 7 M urea, 2 M thiourea, 4% CHAPS, 1 mM EDTA, one tablet of Complete Protease Inhibitor (Roche, Basel; Switzerland) per 100 ml, and one tablet of PhosStop Protease Inhibitor (Roche) per 100 mL). Total protein concentration was determined with the 2D-Quant kit (GE Healthcare, Uppsala, Sweden) per manufacturer’s manual. Lysates were stored at -80°C until further processing. We investigated the molecular mechanisms of endothelial cell pathology during in vivo and in vitro PD fluid exposure by proteomic and bioinformatic analysis of the endothelial cell response to hyperglycemic stress, integrating current understanding of PD-related cellular injury and stress responses (Bender et al., Nephrol Dial Transplant, 2011, doi:10.1093/ndt/gfq484; Kratochwill et al., BioMed research international, 2015, doi:10.1155/2015/628158; Kratochwill et al., J Proteome Res, 2009, doi:10.1021/pr800916s; Lechner et al., J Proteome Res, 2010, doi:10.1021/pr9011574). We then evaluated the effect of AlaGln addition to conventional PD fluid on these parameters in endothelial cells. This strategy allowed characterization of endothelial cell injury and stress responses during exposure to conventional hyperglycemic PD fluid, and their modulation by added AlaGln.
2021-09-09 | PXD022170 | Pride