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We demonstrated that NK cell activity affects HSCs frequency in vitro as well as hematopoietic reconstitution in vivo. Gene expression and cytokine profile assays revealed the potential players of this HSC-NK cell regulation, such as IFNγ. Besides the known role of C/EBPg in NK cell differentiation, we demonstrated that it is also essential for NK-cytokine mediated HSC regulation. Finally, NK cell depletion from murine and human bone marrow favored transplant chimerism.
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The aim of this study is to identify sites occupied by Pdx1 throughout key stages in mouse and human pancreatic development as well as during in vitro differentiation of human ES cells.
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To interrogate the hypoxia response pathways affected by the depletion of CAIX in BC cells, we performed gene expression profiling using SurePrint G3 Human Gene Expression 60K microarrays in MDA-MB-231-shCAIX, MCF7-shCAIX and the respective control cells grown as multicellular spheroids under hypoxic or normoxic conditions. The human breast cancer cell lines MDA-MB-231 and MCF7 were purchased from the European Collection of Cell Cultures (ECACC, UK) and maintained as recommended by the manufacturer. To generate stable CAIX silenced cells, cell lines were transfected with a pool of three plasmids each encoding CAIX-specific shRNA (1 µg) and a negative control (NC) plasmids encoding scrambled shRNAs (1 µg) (Santa Cruz Biotechnology, TX, USA) using shRNA transfection medium and shRNA transfection reagent (Santa Cruz Biotechnology, USA) according to the manufacturers’ protocol. For selection of stably transfected cells, puromycin dihydrochloride (Santa Cruz Biotechnology, USA) was added to medium 48 h post-transfection (6 µg/ml for MCF7, 2 µg/ml for MDA-MB-231 cells). The transfected cells were plated at density 2e+3 cells per ml of serum-free DMEM/F12 medium supplemented with EGF (20ng/ml, R&D Systems, #236-EG-200), bFGF (10ng/ml, SantaCruz, #sc-4573), hydrocortisone (50 ng/ml, Sigma-Aldrich, #H0135-1MG) and 1B27 (Invitrogen, #17504001) and grown as multicellular spheroids for 5 days. To establish hypoxic conditions, the cells were cultured at 1% oxygen, 94% N2 and 5% CO2 at 37oC using humidified multi-gas incubator (Sanyo, Sanyo Electric Co.,Ltd.) for 48 hours. The normoxic control cells were incubated at 37oC with 5% CO2 in a humidified incubator (Panasonic, Panasonic Healthcare Co., Ltd.). The efficiency of CAIX silencing was assessed by qRT-PCR and immunofluorescence before the experiments.
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mRNA expression data in FACS sorted CD11b+ F4/80+ tumor-associated macrophages that are derived from normoxic (as characterized by low GLUT1-expression on the cell membrane) or hypoxic (as characterized by high GLUT1-expression on the cell membrane) area's of Lewis Lung Carcinoma tumors injected subcutaneously in either wild type or REDD1 (DDIT4) knockout mice.
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Aim: To determine the impact of NUAK1 depletion on the transcriptome of U2OS cells.
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It is unclear why preterm birth increases risk of cardiovascular disease later in life. Studies in mice indicate excess oxygen used to treat preterm infants causes pulmonary hypertension, cardiac failure, and shortens lifespan. We previously reported neonatal hyperoxia causes pulmonary hypertension in aged mice as defined pathologically by pulmonary capillary rarefaction, dilation of pulmonary arterioles and veins, right ventricular hypertrophy, and reduced lifespan. Here, affymetrix gene arrays were used to identify early transcriptional changes in lungs of young adult mice exposed to room air or 100% oxygen between postnatal days 0-4.
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621-101 cells were treated with rapamycin and rhebsiRna and samples were run in triplicates for each condition.
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Elavl1/HuR is a ubiquitous and conserved RNA-binding protein that binds to a U-rich RNA motif that shuttles between nucleus and cytoplasm. In epithelia, the elevated expression of HuR assumingly promotes degeneration and cancer suggesting that its generic suppression may provide clinical benefits. In this study we focused on biological and clinical functions of HuR in intestinal epithelial cells and we presented evidence that changes in HuR levels induce polarized distortions in these cells to support different pathologic outcomes. For translation profiling cytoplasmic fractions of CMT93 sublines, containing monosomes and polysomes were separated as in (Katsanou et al., 2009). Fractions containing monosomes and polysomes were collected using a retriever 500 fraction collector (Teledyne Isco). Monosomal and polysomal fractions were pooled and total RNA was extracted from each fraction using Trizol reagent (Invitrogen). For micorarrays, RNAs were further purified via columns (Qiagen). Biotinylated complementary RNAs,(cRNAs) were hybridized onto Affymetrix Gene Mo-Gene-1.0 Chip in accordance to the protocols of the Genomics Unit of BSRC “Alexander Fleming”. (http://www.fleming.gr/facilities/genomics).
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Phoenix dactylifera seedlings were exposed to heat, drought and combined heat & drought conditions in growth chambers. Leaf samples were collected for total RNA isolation (RNAseq, Illumina HiSeq 1000), and water soluble metabolites. The RNAseq of four biological replicates (two individuals per replicate) were compared against the control condition. Transcriptomics data suggests the combine heat and drought resembled heat response, whereas drought resembled more to control. The hallmarks of heat stress were visible in the transcriptomics data, such as protein misfolding, response to hydrogen peroxide and cell wall modification, as well as ABA signaling in the case of drought. Since the plants were exposed to the stress for several days before harvesting, the early signs of heat stress such as calcium and NO signaling were not detected anymore. In addition, data suggest a significant enrichment of circadian rhythm motifs in the differentially expressed genes in heat and combined heat and drought stresses, suggesting new stress avoidance strategies.
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This experiment aimed at characterising the signalling pathways downstream SUCNR1 activation in murine Neural Stem Cells (NSCs). To this end, we profiled by microarray the gene expression changes induced by succinate stimulation in both wild type NSCs and GPR91-deficient NSCs.
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