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Two trophoblastic cell lines, CRL-1584 (3A-Sub E) and JEG-3 were purchased from American Type Culture collection (ATCC) (Manassas, VA). CRL-1584 (3A-Sub E) was originally derived from human term primary placental cells and then immortalized using SV40 virus. JEG-3 is a clonal human choriocarcinoma. The cell lines were serially passaged in complete medium supplemented with each cell line’s respective IC50 concentration of methotrexate until the cell lines were able to proliferate normally. Subsequently, the concentration of methotrexate was increased by one half-log concentration. The process was repeated iteratively until the cells became senescent. At each log concentration of methotrexate resistance, cell line stocks were frozen to establish a separate subline of each cell line. Based on their relative level of resistance to methotrexate, these were designated JEG-3-R7, JEG-3-R6, and JEG-3-R5 (for 10-7 M through 10-5 M methotrexate resistance). Over the same time period (approximately 11 months), we were able to induce only one order of magnitude of resistance in the normal placenta cell line; that line was designated NP-R7. In parallel, the parent cell lines were passaged in normal medium to control for any physiologic changes induced by prolonged culture in vitro.
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This experiment sought to determine whether estrogen could directly activate changes in gene transcription in lung and uterine ILC2s, whether such changes differed between lung and uterine ILC2s, and compared baseline gene expression between ILC2s from those organs. ILC2s, defined as lineage (CD3e, CD14, CD16/32 and B220) negative, CD25 positive and CD44 high lymphocytes, were sorted by flow cytometry from 8-12 week old BALB/c mice and cultured overnight in RPMI 1640 with 10% charcoal/dextran treated calf serum in the absence or presence of 100 ng/ml estrogen. RNA was extracted from the cells after culture.
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Decidual spiral arteriole (SpA) remodelling is essential to ensure optimal uteroplacental blood flow during human pregnancy. Decidual uterine natural killer cells and macrophages infiltrate the SpA and are proposed to initiate remodelling before colonisation by extravillous trophoblasts. Microarrays were used to measure the effect of extravillous trophoblasts conditioned medium on the transcriptome of human uterine microvascular endothelial cells.
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Investigating a mouse model of Cushing syndrome, synthetic adrenocorticotropic hormone ACTH (Synacthen) was infused for two weeks to markedly increase adrenal mass and plasma corticosterone levels. Gene expression in adrenals was measured to look for changes associated with the adrenal cellular phenotype.
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The experiment intended to identify biomarker genes for arsenite exposure in rice roots. These biomarker genes were tested as alternative endpoints in an sediment-contact test with rice (Brinke et al. Environ Sci Pollut R 22.16 (2015): 12664-12675)
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Endothelial colony-forming cells were isolated and expanded from the mononuclear cell fraction (MNC) obtained from the cord blood of term (CT) and preterm (PT) neonates, and characterized as previously described (Vassallo PF et al 2014). The objective of the study was to identify differentially expressed genes between CT and PT neonates. ECFC were harvested from cultures dishes at passage 3 and total RNA was extracted using the mirVana miRNA Isolation Kit (Ambion), according to the manufacturer’s recommendations. Cy3-CTP labeled RNA was prepared according to standard Agilent protocol from 400ng total RNA. The hybridization was performed for 17 hrs at 65°C. cRNA labeling and hybridization performance were performed and all parameters checked were found within the manufacturers specifications. Arrays were scanned as described in the manufacturers’ protocol. Signal intensities on 20 bit tiff images were calculated by Feature Extraction software (FE, Version 8.5; Agilent Technologies). Data analyses were conducted with GeneSpring GX software (Vers.13.1.1; Agilent Technologies)
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We measured global mRNA expression in peripheral blood mononuclear cells (PBMC) from 182 individuals, comprising of 142 multiple sclerosis (MS) patients affected with distinct MS clinical forms and 40 healthy controls.
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We measured global mRNA expression in peripheral blood mononuclear cells (PBMC) from 76 individuals, comprising of 49 multiple sclerosis (MS) patients affected with distinct MS clinical forms and 27 healthy controls.
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We have performed transcriptomic analysis in the spinal cord of experimental autoimmune encephalomyelitis (EAE) mice compared to naive mice at different time intervals in order to observe the gene expression changes within the CNS compartment
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Introduction of PTEN pseudogene in murine breast cancer upregulates p53 and activating protein 2 gamma and delays tumor growth.
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