ArticleBMC molecular biology2010
Effect of experimental treatment on GAPDH mRNA expression as a housekeeping gene in human diploid fibroblasts.
Article in BMC molecular biology, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 70 citations in OpenAlex.
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- Identification of the Reference Genes for Relative qRT-PCR Assay in Two Experimental Models of Rabbit and Horse Subcutaneous ASCs.International journal of molecular sciences · 2024Article
- Identification of Accurate Reference Genes for qRT-PCR Analysis of Gene Expression inPlants (Basel, Switzerland) · 2023Article
- Comprehensive Analysis of Gene Expression Profiling to Explore Predictive Markers for Eradication Therapy Efficacy againstCancers · 2023Article
- Maternal separation in rats induces neurobiological and behavioral changes on the maternal side.Scientific reports · 2020Article
- Chlorella vulgaris modulates the expression of senescence-associated genes in replicative senescence of human diploid fibroblasts.Molecular biology reports · 2020Article
- Selection of suitable reference genes for gene expression studies in myxosporean (Myxozoa, Cnidaria) parasites.Scientific reports · 2019Article
- Cytoplasmic delivery of functional siRNA using pH-Responsive nanoscale hydrogels.International journal of pharmaceutics · 2019Article
- High expression of acidic chitinase and chitin digestibility in the stomach of common marmoset (Callithrix jacchus), an insectivorous nonhuman primate.Scientific reports · 2019Article
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- Chitin digestibility is dependent on feeding behaviors, which determine acidic chitinase mRNA levels in mammalian and poultry stomachs.Scientific reports · 2018Article
- Impaired redox environment modulates cardiogenic and ion-channel gene expression in cardiac-resident and non-resident mesenchymal stem cells.Experimental biology and medicine (Maywood, N.J.) · 2017Article
- γ-Tocotrienol prevents cell cycle arrest in aged human fibroblast cells through p16Journal of physiology and biochemistry · 2017Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundSeveral genes have been used as housekeeping genes and choosing an appropriate reference gene is important for accurate quantitative RNA expression in real time RT-PCR technique. The expression levels of reference genes should remain constant between the cells of different tissues and under different experimental conditions. The purpose of this study was to determine the effect of different experimental treatments on the expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNA so that the reliability of GAPDH as reference gene for quantitative real time RT-PCR in human diploid fibroblasts (HDFs) can be validated. HDFs in 4 different treatment groups viz; young (passage 4), senescent (passage 30), H2O2-induced oxidative stress and gamma-tocotrienol (GTT)-treated groups were harvested for total RNA extraction. Total RNA concentration and purity were determined prior to GAPDH mRNA quantification. Standard curve of GAPDH expression in serial diluted total RNA, melting curve analysis and agarose gel electrophoresis were used to determine the reliability of GAPDH as reference gene.
resultsHDFs with different experimental treatments exhibited diverse cell morphology with different expression of senescence-associated beta-galactosidase (SA beta-gal) activity. However the expression level of GAPDH was consistent in all treatment groups.
conclusionThe study demonstrated that GAPDH is reliable as reference gene for quantitative gene expression analysis in HDFs. Therefore it can be used as housekeeping gene for quantitative real time RT-PCR technique in human diploid fibroblasts particularly in studying cellular senescence.
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