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| Status |
Public on Apr 01, 2012 |
| Title |
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 |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by RT-PCR
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| Summary |
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.
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| Overall design |
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.
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| Contributor(s) |
Honda N |
| Citation(s) |
22379029 |
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| Submission date |
Jan 03, 2012 |
| Last update date |
May 02, 2012 |
| Contact name |
Noritoshi Honda |
| Organization name |
Kumamoto University
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| Department |
Department of Dermatology and Plastic Surgery
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| Street address |
1-1-1 Honjo
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| City |
kumamoto |
| State/province |
kumamoto |
| ZIP/Postal code |
860-8556 |
| Country |
Japan |
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| Platforms (1) |
| GPL15085 |
SABiosciences Human Cell Development & Differentiation miRNA PCR ArrayHuman panel |
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| Samples (3) |
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| Relations |
| BioProject |
PRJNA150233 |
