Browse
Submit Data
Databases
API
Help

Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

16 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in systemic sclerosis (SSc) dermal fibroblasts

ABSTRACT: Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs. Human dermal fibroblasts were obtained by skin biopsy from the affected areas (dorsal forearm) of 5 patients with diffuse cutaneous SSc and <2 years of skin thickening. Control fibroblasts were obtained by skin biopsies from 5 healthy donors. Control donors were each matched with a SSc patient for age, sex, and biopsy site.

ORGANISM(S): Homo sapiens

SUBMITTER: Taiji Naashima

PROVIDER: E-GEOD-34142 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in normal fibroblasts stimulated with exogenous TGF-β1 and systemic sclerosis (SSc) dermal fibroblasts

Project description:Normal fibroblasts and SSc fibroblasts between the third and six subpassages were used for experiments. Normal and scleroderma fibroblasts were serum-starved for 24 hours and incubated in the presence or absence of TGF-β1 (2ng/ml) for 6 hours. Total RNA was extracted from culture cells with ISOGEN (Nippon Gene, Tokyo, Japan). MicroRNA isolation from total RNA was performed using RT2 qPCR-Grade miRNA Isolation Kit (SA Bioscience). For RT2 Profiler PCR Array (SABioscience), microRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of cDNAs from 5 normal fibroblasts or 5 SSc fibroblasts was prepared. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into a 96-well RT2 miRNA PCR Array (SABiosciences) that included primer pairs for 88 human microRNAs. Human dermal fibroblasts were obtained by skin biopsy from the affected areas (dorsal forearm) of 5 patients with diffuse cutaneous SSc and <2 years of skin thickening. Control fibroblasts were obtained by skin biopsies from 5 healthy donors. Control donors were each matched with a SSc patient for age, sex, and biopsy site.

2012-04-01 | E-GEOD-34827 | biostudies-arrayexpress

Real-time quantitative PCR analysis of 88 microRNAs involved in human cell differentiation and development in involved skin of systemic scleroderma (SSc), localized scleroderma (LSc) and Keloid.

Project description:Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. The microRNA (miRNA) isolation from human skin tissue was performed using miRNeasy FFPE kit (Qiagen). For PCR array, miRNAs were reverse-transcribed into first strand cDNA using RT2 miRNA First Strand Kit (SABiosciences). A mixture of equal amounts of miRNAs from 3 normal skins, 3 SSc, 3 LSc or 3 keloid were prepared, and miRNA expression profile in each disease in vivo was evaluated using RT2 Profiler PCR Array. The cDNA was mixed with RT2 SYBR Green/ROX qPCR Master Mix and the mixture was added into 96-well RT2 miRNA PCR Array that includes primer pairs for 88 human miRNAs (SABiosciences). Skin specimens were derived from involved skin of 3 systemic scleroderma (SSc), 3 localized scleroderma (LSc) and 3 keloid patients. These skin samples and 3 control skins were collected and fixed in formaldehyde immediately after resections. Control donors were each matched with diseases for age, sex, and biopsy site.

2013-01-15 | E-GEOD-43469 | biostudies-arrayexpress

Profiling Period 2 (PER2) mediated pathways following ischemic preconditioning (IPC) of the mouse heart using a genome array with 352 abundantly expressed and well-characterized micro RNAs

Project description:Ischemic preconditioning (4 cycles of 5 min ischemia and 5 min reperfusion) with a final reperfusion time of 120 minutes was performed in C57BL/6N (The Jackson Laboratory) or Per2-/- mice. Heart tissue was snap-frozen with clamps pre-cooled to the temperature of liquid nitrogen. Micro RNA was isolated with Trizol (Invitrogen) and purified using RT2 qPCR-Grade miRNA Isolation Kit (SABiosciences-Qiagen). cDNA template was generated using RT2 miRNA First Strand Kit (SABiosciences-Qiagen). miRNA expression was performed using RT2 miRNA PCR Array Mouse miFinder (SABiosciences-Qiagen).

2017-04-12 | E-MTAB-5401 | biostudies-arrayexpress

miRNA profiling in hPASMC isolated from normal and PAH patients

Project description:To identify dysregulated miRNAs in PAH HPASMC, we compared the miRNA expression profiles between normal and PAH HPASMC using the RT2 miRNA PCR Array System. Total RNA was isolated using a miRNeasy Mini Kit (Qiagen, Valencia, CA) and treated with an RNase-Free DNase Set (Qiagen). After quantification with Nanodrop 2000 spectrophotometer (ThermoScientific, Rockford, IL), miRNAs were reversely transcribed using a RT2 miRNA First Strand Kit (SABiosciences, Frederick, MD). The RT2 miRNA PCR Array System (SABiosciences) was used to study miRNA profiling in normal and PAH HPASMC. U6, SNORD44, SNORD47 and SNORD48 were used as internal controls for the profiling. HPASMC isolated from oe normal donor (C128, as control), one IPAH patient (B157), one PAH patient with Eisenmenger’s syndrome (CC-008) and one PAH patient with Scleroderma (CCF-004) were used for miRNA PCR array study. Data is normalized to the control sample. Three independent sample preparations were used for this study.

2014-02-28 | E-GEOD-55427 | biostudies-arrayexpress

The expression profiles of extracellular matrix-related genes in the presence or absence of IL-17A or -17F as measured with the PCR array in systemic sclerosis (SSc) dermal fibroblasts

Project description:We examined the effect of IL-17 signaling pathway on extracellular matrix (ECM) expression and the involvement of IL-17 signaling pathway in pathogenesis of SSc. To identify differences in the expression pattern of ECM genes in IL-17A- or IL-17F-treated cells, we performed PCR array analysis, consisting of 84 ECM-related genes. Normal human dermal fibroblasts were cultured until they were confluent, and then stimulated with IL-17A or IL-17F for 12 hours, and total RNA was extracted. A mixture of equal amounts of mRNAs from three normal fibroblasts was prepared in the presence or absence of IL-17A or -17F, and mRNA expression profile was evaluated using PCR Array. Normal fibroblasts were obtained by skin biopsies from 3 healthy donors. Fibroblasts from donors were used and treated separately as indicated in the summary. Equal amount total RNA from each donor was pooled prior to gene expression analysis.

2012-01-01 | E-GEOD-33581 | biostudies-arrayexpress

Real-time quantitative PCR analysis of the expression of chromatin modification enzyme genes in 4 week-old male MeCP2 A140V mutant mouse cerebellum

Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates.The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A) from SABiosciences. Mice used in this study were 4 week old males. Total RNA was extracted from whole cerebella of 4 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A).

2015-02-01 | E-GEOD-35689 | biostudies-arrayexpress

Real-time quantitative PCR analysis of the expression of chromatin modification enzyme genes in 2 week-old male MeCP2 A140V mutant mouse cerebellum

Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from whole cerebella of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Epigenetic Chromatin Modification Enzymes Array (Catalog# PAMM-085A).

2015-02-01 | E-GEOD-35688 | biostudies-arrayexpress

Real-time quantitative PCR analysis of the expression of cytoskeleton regulator genes in 2 week old male MeCP2 A140V mutant mouse brain cortex

Project description:The goal of this study was to identify brain cortex gene expression differences between mutant MeCP2 A140V mice and their wild type littermates.The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from the somatosensory/motor cortex of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cortical tissue from 3 mice of each genotype was used and RNA from the cortex of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A).

2015-02-01 | E-GEOD-35769 | biostudies-arrayexpress

Real-time quantitative PCR analysis of the expression of cytoskeleton regulator genes in 4 week old male MeCP2 A140V mutant mouse cerebellum

Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates.The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A) from SABiosciences. Mice used in this study were 4 week-old males. Total RNA was extracted from whole cerebella of 4 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A).

2015-02-01 | E-GEOD-35691 | biostudies-arrayexpress

Real-time quantitative PCR analysis of the expression of cytoskeleton regulator genes in 2 week old male MeCP2 A140V mutant mouse cerebellum

Project description:The goal of this study was to identify cerebellar gene expression differences between mutant MeCP2 A140V mice and their wild type littermates. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A) from SABiosciences. Mice used in this study were 2 week old males. Total RNA was extracted from whole cerebella of 2 week old male MeCP2 A140V hemizygous mutant mice and their wild type littermates. For this study, cerebella from 3 mice of each genotype were used and RNA from the cerebellum of each mouse was analyzed separately (ie., triplicate biological replicates). The RNA was purified to remove genomic DNA and was then reverse transcribed for use in quantitative RT-PCR analysis of gene expression using the SABiosciences RT2 profiler PCR array system. The PCR array used for these experiments was the Mouse Cytoskeleton Regulators Array (Catalog# PAMM-088A).

2015-02-01 | E-GEOD-35690 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data