Notes & TipsRapid detection of CSRP2 mRNA in mouse, rat, and human using LightCycler-based quantitative real-time polymerase chain reaction☆
Section snippets
Cells, tissue samples, RNA isolation, Northern blot
Total cellular RNAs from mouse tissue samples were isolated by a standard procedure [19]. Isolation of primary rat hepatic stellate cells and culturing of human cell line WI-26 VA4 were performed as described previously [8], [20], [21]. For Northern blot analysis RNAs were separated in a 1.2% denaturing agarose gel, transferred to nylon, and probed with a -labeled EcoRI fragment of clone pCRIII-rSmLIM encoding rat CSRP2[7] or a β-actin-specific cDNA probe.
cDNA preparation and PCR
Total RNA (2 μg) was reverse
Results and discussion
By use of traditional techniques (e.g., Northern blot, in situ hybridization) CSRP2 expression was found in various tissues of mammals. However, these methods are only semiquantitative and in view of the fact that suppression of CSRP2 is correlated with cellular transformation or in conjunction with inflammatory and fibroproliferative responses there is growing interest in the development of methods permitting one to monitor the transcription level of CSRP2 more precisely. Over the past years,
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Molecular cloning, characterization and tissue specificity of the expression of the ovine CSRP2 and CSRP3 genes from Small-tail Han sheep (Ovis aries)
2016, GeneCitation Excerpt :Additionally, phylogenetic analysis indicated that mammalian CRP2 or CRP3 have clearly recognizable orthologs in lower vertebrates, suggesting that certain CRP2 and CRP3 have been evolved before species divergence. CSRP2 mRNA showed remarkably tissue-specific expression in the blood vessels, which is consistent with the pattern in mice (Günther et al., 2003) and pigs (Xu et al., 2010). CSRP2 mRNA expression in the aorta was greater in DP than in SH (P < 0.05).
Caffeine suppresses the expression of the Bcl-2 mRNA in BeWo cell culture and rat placenta
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This work was supported by Deutsche Forschungsgemeinschaft Grant We2554/1 to R.W.