RGD Reference Report - Quantitative monitoring of the mRNA expression pattern of the TGF-beta-isoforms (beta 1, beta 2, beta 3) during transdifferentiation of hepatic stellate cells using a newly developed real-time SYBR Green PCR. - Rat Genome Database

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Quantitative monitoring of the mRNA expression pattern of the TGF-beta-isoforms (beta 1, beta 2, beta 3) during transdifferentiation of hepatic stellate cells using a newly developed real-time SYBR Green PCR.

Authors: Wickert, L  Steinkruger, S  Abiaka, M  Bolkenius, U  Purps, O  Schnabel, C  Gressner, AM 
Citation: Wickert L, etal., Biochem Biophys Res Commun 2002 Jul 12;295(2):330-5.
RGD ID: 730145
Pubmed: PMID:12150952   (View Abstract at PubMed)

Current methods to determine the mRNA of the TGF-beta-isoforms, beta 1, beta 2, and beta 3, are not sensitive enough to detect small alterations in the expression levels. Therefore, we established a SYBR Green I-based real-time quantitative PCR procedure with fragment-specific standards. The advantage of gene-specific quantification is the possibility to be abstain from the need to compare results with a house-keeping gene having a different sequence and PCR efficiency. Reproducibility of the results and analytical variances of the real-time PCR assays were tested. In transdifferentiating rat hepatic stellate cells (HSC) the TGF-beta 1-mRNA was found to be the predominant isoform expressed followed by TGF-beta 3 and low amounts of TGF-beta 2-mRNA. An alteration of the TGF-beta 1,-beta 2, and -beta 3 ratio during HSC transdifferentiation could not be detected. Furthermore, the GAPDH mRNA expression varied during HSC activation, and thus is not recommended as a standard in real-time PCR quantifications.

Objects referenced in this article
Gene Tgfb1 transforming growth factor, beta 1 Rattus norvegicus
Gene Tgfb2 transforming growth factor, beta 2 Rattus norvegicus
Gene Tgfb3 transforming growth factor, beta 3 Rattus norvegicus

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