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RNA sequencing of extracellular vesicles, secreted by young and adult mouse adipocytes [miRNA-Seq]

ABSTRACT: We used male C57BL/6 mice (Charles River Laboratories, Wilmington, MA), housed under SPF conditions. Primary adipocytes from the inguinal fat depot at postnatal day (P6, young mice) and postnatal day 56 (P56, adult mice) were isolated by collagenase digestion and separation of cell fractions and subsequently cultured in EV-free media (FBS was EV-depleted). Article: Röszer T., MicroRNA Profile of Mouse Adipocyte-Derived Extracellular Vesicles

ORGANISM(S): Mus musculus

PROVIDER: GSE193589 | GEO | 2024/08/01

REPOSITORIES: GEO

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RNA sequencing of extracellular vesicles, secreted by young and adult mouse adipocytes [RNA-Seq]

Project description:We used male C57BL/6 mice (Charles River Laboratories, Wilmington, MA), housed under SPF conditions. Primary adipocytes from the inguinal fat depot at postnatal day (P6, young mice) and postnatal day 56 (P56, adult mice) were isolated by collagenase digestion and separation of cell fractions and subsequently cultured in EV-free media (FBS was EV-depleted). Article: Röszer T., MicroRNA Profile of Mouse Adipocyte-Derived Extracellular Vesicles

2024-08-01 | GSE191256 | GEO

Transcription factors expressed in olfactory bulb local progenitor cells revealed by genome-wide transcriptome profiling

Project description:We used transcriptional profiling to monitor gene expression of whole olfactory bulbs at daily intervals from embryonic day 11 through birth, generating a compendium of gene expression encompassing the major developmental events of this tissue. OBs from CD1 mice (Charles River Laboratories, Wilmington, MA), were isolated from embryos daily between E11 and postnatal day zero (P0), for a total of nine time points. RNA was purified with TRIzol reagent (Invitrogen, Carlsbad, CA), subjected to two rounds of amplification, labeled, and hybridized to Affymetrix Mouse Genome 430.2 GeneChip microarrays (Affymetrix Inc., Santa Clara, CA, USA) using Affymetrix reagents and protocols (http://www.affymetrix.com). One microarray for each time point using RNA from the OBs of one individual embryo.

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Sex and Molecular Differences in Cardiovascular Parameters at Peak Influenza Infection in Mice

Project description:Female and male C57BL/6J mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA) at 16 weeks age. Mice were intranasal infected with a sublethal does of 7.5×104 EID50 A/Puerto Rico/8/34 influenza virus (PR8; Charles River, Wilmington, MA, USA) or mock infected (naïve) with 1X PBS. After euthanasia at 7 days post infection lung and nasal septum were collected from animals to perform transcriptome analysis by RNA sequencing.

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Gene expression analysis of mouse adipose tissue macrophages

Project description:We collected adipose tissue macrophages from C57Black/6 mice on postnatal day 6 (P6) and postnatal day 56 (P56) by enzyntic digestion of the inguinal fat depots and megnetic cell sorting of F4/80 antigen positive macrophages. Total RNA was isolated and used for next generation RNA sequencing.

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A murine pneumonic plague model after infection with wild-type Yersinia pestis CO92 and its Braun lipoprotein mutant

Project description:Swiss-Webster female mice (Charles River Laboratories, Wilmington, MA) 5-6 weeks of age were infected intranasally with 5 LD50 of either WT or Δlpp mutant of Y. pestis CO92. Uninfected mice were used as controls. At either 12 or 48 h post infection (p.i.), 3 mice per group were euthanized and the lungs, livers, and spleens were harvested and homogenized in 1 ml of RNALater (Ambion/Applied Biosystems, Austin, TX) using 50-ml tissue homogenizers (Kendell, Mansfield, MA). RNA was isolated from the tissue homogenates and purified using RNAqueous (Ambion). After an overnight precipitation, the RNA was resuspended in 20 ul of diethylpyrocarbonate (DEPC)-treated water and hybridized to Affymetrix GeneChip Mouse Genome 430 2.0 arrays, performed by the Molecular Genomics Core at UTMB Galveston, Texas, per manufacture protocols. The arrays had 45,000 probe sets representing more than 39,000 transcripts derived from ~34,000 well-substantiated mouse genes. The experiments were performed in triplicate (biological replicates), generating a total of 45 arrays.

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RNA-seq analysis of neonatal mouse cochlear supporting cells [Treated AK]

Project description:This study examined transcripts that are enriched in neonatal mouse cochlear supporting cells at postnatal day 1 and postnatal day 6 after inhibition of the Notch signaling pathway. Cochleas from postnatal day 0 and postnatal day 5 were cultured for 24 hours in the gamma secretase inhibitor DAPT or DMSO as a vehicle control. Supporting cells were purified by FACS sorting for GFP fluorescence from the cochleas of transgenic mice in which a BAC including the LFng locus drives the expression of GFP. Two replicates of GFP+ supporting cells were compared with all other cochlear cell types that were GFP-. We performed this experiment at two different ages, postnatal day 0+24 hours culture and postnatal day 5 + 24 hours culture. (corresponding to P1 and P6).

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RNA-seq analysis of neonatal mouse cochlear supporting cells [NonTreated_JM]

Project description:This study examined transcripts that are enriched in neonatal mouse cochlear supporting cells at postnatal day 1 and postnatal day 6. Supporting cells were purified by FACS sorting for GFP fluorescence from the cochleas of transgenic mice in which a BAC including the LFng locus drives the expression of GFP. Two replicates of GFP+ supporting cells were compared with all other cochlear cell types that were GFP-. We performed this experiment at two different ages, postnatal day 1 and postnatal day 6. mRNA profiles of supporting cells (GFP+) and all other cochlear cell types (GFP-), two replicates each, at P1 and P6 mice were generated by deep sequencing using Illumna TruSeq.

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Mouse liver sinusoidal endothelial cell responses to the glucocorticoid receptor agonist dexamethasone

Project description:Liver sinusoidal endothelial cells (LSECs) from C57Bl/6J male mice obtained from Charles River Laboratory (Sulzfeld, Germany) were cultured on fibronectin coated plates in 5% oxygen atmosphere in AIM-V with serum-free supplements for 1h, 10h, or 48h. Data from quantitative proteomics using a tandem mass tag (TMT)6-plex strategy on an Orbitrap Fusion Lumos mass spectrometer (Thermo Fisher Scientific)), using a TMT SPS MS3 method (Navarrete-Perea, Yu et al. 2018).

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Hedgehog activation promotes osteogenic fates of growth plate resting zone chondrocytes through transient clonal competency [NovaA-186]

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2023-10-11 | GSE244880 | GEO

5 Day Oral Study of A-998679 in Male Sprague Dawley Rats (liver)

Project description:Male Sprague-Dawley rats [Crl:CD®(SD)IGS BR], weighing ~250 g at study initiation were obtained from Charles River Laboratories, Inc. (Wilmington, MA). Rats were housed singly in ventilated, stainless steel, wire-bottom hanging cages and fed non-certified Rodent Chow (Harlan Labs, Madison, WI) and water ad libitum and acclimated for at least 5 days after arrival. Rats were randomly assigned to various treatment groups (3 rats/group) and were dosed once daily by oral gavage with vehicle (0.2% hydroxypropylmethylcellulose at a dose volume of 10 ml/kg) or with 30, 100, or 200 mg/kg of A-998679. All rats were fasted overnight after their last dose, weighed and sacrificed under isoflurane anesthesia. Liver and small intestine (jejunum) were flash frozen in liquid nitrogen and stored at −80°C until processing for gene expression profiling on the Affymetrix platform.

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