title: Selection of reference genes for qRT-PCR in high fat diet-induced hepatic steatosis mice model.
abstract: With the epidemic proportions of obesity worldwide and the concurrent prevalence of hepatic steatosis, there is an urgent need for better understanding the intrinsic mechanism of hepatic steatosis, especially the changes of gene expression underlying the development of hepatic steatosis and its associated abnormal liver function. Quantitative real-time PCR (qRT-PCR) is a sensitive and highly reproducible technique of gene expression analysis. However, for accurate and reliable gene expression results, it is vital to have an internal control gene expressed at constant levels under all the experimental conditions being analyzed for. In this study, the authors validated candidate reference genes suitable for qRT-PCR profiling experiments using livers from control mice and high fat diet-induced obese mice. Cross-validation of expression stability of ten selected reference genes using three popular algorithms, GeNorm, NormFinder, and BestKeeper found HPRT1 and GAPDH as most stable reference genes. Thus, HPRT1 and GAPDH are recommended as stable reference genes most suitable for gene expression studies in the development of hepatic steatosis.
authors.affiliation: Laboratory of Endocrinology and Metabolism, Institute of Health Sciences, Shanghai Institutes for Biological Sciences (SIBS), Chinese Academy of Sciences (CAS) & Shanghai Jiao Tong University School of Medicine (SJTUSM), 225 South Chongqing Road, Shanghai 200025, China.
authors.forename: Lingyan
authors.initials: L
authors.lastname: Xu
authors.forename: Xinran
authors.initials: X
authors.lastname: Ma
authors.forename: Bin
authors.initials: B
authors.lastname: Cui
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Li
authors.forename: Guang
authors.initials: G
authors.lastname: Ning
authors.forename: Shu
authors.initials: S
authors.lastname: Wang
mesh_terms: D000818:Animals; D004041:Dietary Fats / Q000008:administration & dosage; D005234:Fatty Liver / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D020869:Gene Expression Profiling / Q000592:standards*; D005819:Genetic Markers / Q000235:genetics*; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating) / Q000235:genetics; D006651:Histocytochemistry; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D012015:Reference Standards; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods / Q000592:standards*
chemical_list: D004041:Dietary Fats; D005819:Genetic Markers; D024581:Glyceraldehyde-3-Phosphate Dehydrogenase (Phosphorylating); D007041:Hypoxanthine Phosphoribosyltransferase