ArticlePloS one2013
Validation of reliable reference genes for real-time PCR in human umbilical vein endothelial cells on substrates with different stiffness.
Article in PloS one, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 35 citations in OpenAlex.
- Non-invasive and probeless rapid in-vitro monitoring and quantification of HUVECs counts based on FFT impedimetery.BioImpacts : BI · 2024Article
- Matrix stiffness, endothelial dysfunction and atherosclerosis.Molecular biology reports · 2023Review
- Matrix stiffness regulates tumor cell intravasation through expression and ESRP1-mediated alternative splicing of MENA.Cell reports · 2023Article
- A Strategy for the Selection of RT-qPCR Reference Genes Based on Publicly Available Transcriptomic Datasets.Biomedicines · 2023Article
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- Reference Gene Validation in the Brain Regions of Young Rats after Pentylenetetrazole-Induced Seizures.Biomedicines · 2020Article
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- The role of extracellular matrix stiffness in regulating cytoskeletal remodeling via vinculin in synthetic smooth muscle cells.Biochemical and biophysical research communications · 2019Article
- Ionic silicon improves endothelial cells' survival under toxic oxidative stress by overexpressing angiogenic markers and antioxidant enzymes.Journal of tissue engineering and regenerative medicine · 2018Article
- Selection of Reference Genes for Studies of Human Retinal Endothelial Cell Gene Expression by Reverse Transcription-Quantitative Real-Time Polymerase Chain Reaction.Gene reports · 2018Article
- Identification and evaluation of reference genes for qRT-PCR studies in Lentinula edodes.PloS one · 2018Article
- Bacterial Cellulose Shifts Transcriptome and Proteome of Cultured Endothelial Cells Towards Native Differentiation.Molecular & cellular proteomics : MCP · 2017Article
- Evaluation of candidate reference genes for RT-qPCR studies in three metabolism related tissues of mice after caloric restriction.Scientific reports · 2016Article
- Biodegradable Polymers Influence the Effect of Atorvastatin on Human Coronary Artery Cells.International journal of molecular sciences · 2016Article
- Identification of Reference Genes for Quantitative Real Time PCR Assays in Aortic Tissue of Syrian Hamsters with Bicuspid Aortic Valve.PloS one · 2016Article
- Genome-wide identification and characterization of reference genes with different transcript abundances for Streptomyces coelicolor.Scientific reports · 2015Article
- Identification of stable reference genes for gene expression analysis of three-dimensional cultivated human bone marrow-derived mesenchymal stromal cells for bone tissue engineering.Tissue engineering. Part C, Methods · 2015Article
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- Evaluation and selection of candidate reference genes for normalization of quantitative RT-PCR in Withania somnifera (L.) Dunal.PloS one · 2015Article
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Authors and funding
8 authors at 2 institutions in 1 country.
Funding
Abstract
backgroundThe mechanical properties of cellular microenvironments play important roles in regulating cellular functions. Studies of the molecular response of endothelial cells to alterations in substrate stiffness could shed new light on the development of cardiovascular disease. Quantitative real-time PCR is a current technique that is widely used in gene expression assessment, and its accuracy is highly dependent upon the selection of appropriate reference genes for gene expression normalization. This study aimed to evaluate and identify optimal reference genes for use in studies of the response of endothelial cells to alterations in substrate stiffness. METHODOLOGY/PRINCIPAL
findingsFour algorithms, GeNorm(PLUS), NormFinder, BestKeeper, and the Comparative ΔCt method, were employed to evaluate the expression of nine candidate genes. We observed that the stability of potential reference genes varied significantly in human umbilical vein endothelial cells on substrates with different stiffness. B2M, HPRT-1, and YWHAZ are suitable for normalization in this experimental setting. Meanwhile, we normalized the expression of YAP and CTGF using various reference genes and demonstrated that the relative quantification varied according to the reference genes. CONCLUSION/SIGNIFICANCE: Consequently, our data show for the first time that B2M, HPRT-1, and YWHAZ are a set of stably expressed reference genes for accurate gene expression normalization in studies exploring the effect of subendothelial matrix stiffening on endothelial cell function. We furthermore caution against the use of GAPDH and ACTB for gene expression normalization in this experimental setting because of the low expression stability in this study.
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