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Human transthyretin (TTR) gene expression is markedly increased in repair phenotype of hereditary ATTR Schwann cell model

ABSTRACT: TTR is markedly expressed in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (S-DMEM)-incubated TgS1 cells compared with Schwann cell medium (SCM)-incubated cells. c-Jun, Gdnf, and Sox2 were upregulated in S-DMEM-incubated TgS1 cell compared with SCM-incubated cells, while Mpz is suppressed in them.

ORGANISM(S): Mus musculus

PROVIDER: GSE213537 | GEO | 2022/09/23

REPOSITORIES: GEO

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Cadmium exposure induces SNAIL1 expression via downregulation of miR-30e in human bronchial epithelial cells

Project description:Purpose: To understand the molecular mechanisms underlying Cd exposure-induced diseases. Methods: Immortalized human bronchial epithelial cells (BEAS-2B) were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Cellgro) supplemented with 1% Penicillin Streptomycin and 10% Fetal Bovine Serum (FBS, Atlanta Biologicals) at 37 degree C and 5 % CO2. For Cd exposure, 0, 2.5, 5 and 10 µM of CdCl2 was added to the media and the cells were cultured for 72h. Results: This study shows that cadmium exposure induces SNAIL1 expression via miR-30e downregulation and the cells undergo epithelial-mesenchymal transition.

2019-06-16 | GSE128263 | GEO

DNA methylation analysis of mouse embryonic fibroblasts (MEFs) and Thymidine treatment-induced MEFs (induced cells).

Project description:Investigation of DNA methlation changing in thymidine treatment-induced MEFs. Mouse embryonic fibroblasts (MEFs) were obtained at E14.5 embryos and supplied with Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) with 15% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. Potential hematopoietic progenitor cells (CD45+CD41+CD31+c-Kit+) were negatively selected with Biotin labeled anti-CD45, anti-CD31, anti-CD41 and anti-c-Kit antibodies by magnetic beads. Hereafter, treated MEFs are referred to as 4neg MEFs for initiation of induction. 4neg MEFs were seeded on mitomycin (MMC)-treated MEFs as feeder cells and cultured in DMEM with 15% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 100 μM thymidine as induction medium for 2 days followed with analysis

2020-01-28 | GSE144272 | GEO

Expression analysis of mouse embryonic fibroblasts (MEFs) and Thymidine treatment-induced MEFs (induced cells)

Project description:Investigation of differentially expressed genes in thymidine treatment-induced MEFs. Mouse embryonic fibroblasts (MEFs) were obtained at E14.5 embryos and supplied with Dulbecco’s modified Eagle’s medium (DMEM) (Invitrogen) with 15% FBS, 100 μg/ml streptomycin and 100 U/ml penicillin. Potential hematopoietic progenitor cells (CD45+CD41+CD31+c-Kit+) were negatively selected with Biotin labeled anti-CD45, anti-CD31, anti-CD41 and anti-c-Kit antibodies by magnetic beads. Hereafter, treated MEFs are referred to as 4neg MEFs for initiation of induction. 4neg MEFs were seeded on mitomycin (MMC)-treated MEFs as feeder cells and cultured in DMEM with 15% FBS, 100 μg/ml streptomycin, 100 U/ml penicillin, 2 mM L-glutamine, 0.1 mM non-essential amino acids and 100 μM thymidine as induction medium for 6 days. Cells were then trypsinized and re-seeded to new feeder cells for another 6 days. Cells were positively selected with Biotin labeled anti-CD45 by magnetic beads followed with analysis.

2020-01-11 | GSE143418 | GEO

Transcriptomic analysis on the effects of melatonin in gastrointestinal carcinomas

Project description:The gastric adenocarcinoma cell line HGC-27 and colorectal adenocarcinoma cell line HCT-8 cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (Hyclone, Logan, UT, USA.). The human hepatocellular carcinoma cell lines HepG2 and Huh-7 were cultured in Dulbecco’s modified Eagle's medium (DMEM) (Hyclone, Logan, UT, USA.).

2019-06-04 | GSE132119 | GEO

Primary muscle cells from in-utero undernourished mice

Project description:ICR mice, age 6-8 weeks.(Harlan, Indianapolis, IN, USA). Housed individually with controlled temperature, humidity, and light-dark cycle (0600h – 1800h). Pregnant mice were housed individually with ad libitum access to standard rodent chow (T.2018, Harlan Teklad, Indianapolis, IN, USA). On day 12.5 of pregnancy, dams were randomly assigned to either a control or undernutrition group. In the undernutrition group, food was restricted to 50% that of gestational day -matched controls for the remainder of pregnancy. After delivery, mothers were given ad libitum access to chow and 24 hours after birth, litters were equalized to eight. We studied mice from 2 groups: in utero undernourished offpring (U) and control offspring (C). Primary muscle cells were isolated from the quadriceps of U and C at 3 weeks old. Isolated primary muscle cells were grown on Matrigel coated flasks in low glucose (5.5 mmol/L) Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 20% FBS, 1X antibiotic-antimycotic, 2.5μg/ml gentamycin and 2.5ng/ml basic fibroblast growth factor. When cells reached approximately 90% confluency, they were differentiated in low glucose DMEM supplemented with 2% FBS, 1X antibiotic-antimycotic, and 2.5μg/ml gentamycin. Other cells were cultured overnight in substrate limited medium (glucose and glutamine free DMEM supplemented with 0.5mM glucose, 1mM glutamine, 0.5mM carnitine and 1% (v/v) FBS; pH 7.4).

2019-09-12 | GSE103689 | GEO

RNAseq of WFU3 mouse prostate epithelial stem cells with knockdown of Map3k7, Chd1, and both

Project description:Purpose: The goal of this study is to identify transcriptome-wide changes that occur in a mouse prostate cell line with knockdown of Map3k7(Tak1) and Chd1. We utilize this cell line as a model of an aggressive prostate cancer subtype consisting of loss of both MAP3K7 and CHD1 in human patients. RNAseq comparing shControl, shMap3k7(Tak1), shChd1, and shDouble (both) was used to identify transcriptome changes resulting from loss of these genes of interest in order to identify potential therapeutic vulnerabilites. Cell line: The generation of the WFU3 cell line has been previously described (Barclay, W.W et al. The Prostate. 2005; PMID:15599944). Breifly, anterior prostate lobes of adult B1/6; 129/SVEV mice were harvested, minced, and extensively digested, and resulting organoids were separated from stromal cells using a Percoll density gradient. These organoids were then plated on rat-tail collagen coated plates in complete growth media, and after 4-5 passages were transferred to uncoated plates. These prostatic epithelial progenitor stem cells are maintained in supplemented 50:50 Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F-12 medium (F12). Complete media components are described in the Barclay, W.W paper.

2019-07-01 | GSE132664 | GEO

Human Sumoylome (SUMO-2) upon bacterial infection

Project description:Given the role of SUMOylation in pathogenic infection, we wanted to evaluate the changes to the host SUMO-2 subproteome during Shigella infection. HeLa cells overexpressing TAP-SUMO-2 or TAP-empty were used in conjunction with SILAC (Stable Isotope Labeling of Amino acids in Culture) (Golebiowski et al, 2009, 2010). Cells were grown in Dulbecco’s modified Eagle’s medium except that L-arginine and L-lysine were replaced with stable isotope (SILAC) forms depending on the treatment. Medium was supplemented with 10% dialyzed fetal calf serum. SILAC experiment compared TAP-containing cells (Lys0 and Arg0) with invasive Shigella flexneri (M90T) infected TAP-SUMO-2-containing cells (4,4,5,5-D4-lysine, Lys4, and 13C6- arginine, Arg6) and TAP-SUMO-2-containing cells infected with (mxiD) non-invasive strain of Shigella (13C6 15N2-lysine, Lys8, and 13C6 15N4 -arginine, Arg10).

2018-10-19 | PXD001100 | Pride

Real-time quantitative PCR analysis of UPR genes in human normal and osteoarthritic chondrocytes

Project description:Human chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from individuals within 12 hours of death (normal) and from patients undergoing total knee arthroplasty (osteoarthritic, OA). The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. Total RNA was extracted from the primary chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73749 | GEO

Real-time quantitative PCR analysis of UPR genes in human osteoarthritic chondrocytes specimens treated with thapsigargin compared to the untreated (control) chondrocytes

Project description:Human osteoarthritic (OA) chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from three patients undergoing total knee arthroplasty. The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. The chondrocytes from each of the three donors were left untreated (control) and treated with thapsigargin (50nM for 20hours).Total RNA was extracted from the chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73746 | GEO

Real-time quantitative PCR analysis of UPR genes in human osteoarthritic chondrocytes specimens treated with tunicamycin compared to the untreated (control) chondrocytes

Project description:Human osteoarthritic (OA) chondrocytes were obtained from the cartilage of femoral condyles and tibial plateaus from three patients undergoing total knee arthroplasty. The chondrocytes were released from the cartilage, seeded at high density, and cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% heat-inactivated fetal bovine serum and an antibiotic mixture (100 units/ml penicillin base and 100 μg/ml streptomycin base) at 37°C in a humidified atmosphere of 5% CO2/95% air. The chondrocytes from each of the three donors were left untreated (control) and treated with tunicamycin (500ng/ml for 20 hours).Total RNA was extracted from the chondrocytes and processed for qPCR according to the manufacturer's instructions (SA Biosciences RT2 Profiler PCR Array PAHS-089Z).

2015-10-06 | GSE73748 | GEO

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