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Housekeeping gene expression variability in differentiating and non-differentiating 3T3-L1 cells
Housekeeping gene expression variability in differentiating and non-differentiating 3T3-L1 cells
Danang Dwi Cahyadi
(Joint Graduate School of Veterinary Sciences, Tottori University, Tottori, Tottori, Japan)
Tomoko Warita
(Department of Biomedical Sciences, School of Biological and Environmental Sciences, Kwansei Gakuin University, Sanda, Hyogo, Japan)
Nanami Irie
(Graduate School of Science and Technology, Kwansei Gakuin University, Sanda, Hyogo, Japan)
Kana Mizoguchi
(Graduate School of Science and Technology, Kwansei Gakuin University, Sanda, Hyogo, Japan)
Jiro Tashiro
(Joint Graduate School of Veterinary Sciences, Tottori University, Tottori, Tottori, Japan)
Yoshinao Z. Hosaka
(Department of Bioresource Sciences, Faculty of Agriculture, Kyushu University, Fukuoka, Fukuoka, Japan)
Katsuhiko Warita
(Joint Graduate School of Veterinary Sciences, Tottori University, Tottori, Tottori, Japan)
Journal: Adipocyte
•
• Taylor & Francis Group
• ISSN: 2162-397X
Abstract
ABSTRACTNormalization is a crucial step in gene expression analysis to avoid misinterpretation. Reverse transcription-quantitative polymerase chain reaction was used to measure the expression of 10 candidate housekeeping genes in non-differentiated (ND) and differentiated (DI) 3T3-L1 cells on days 5 and 10. We used geNorm, NormFinder, BestKeeper, RefFinder, and the ∆Ct method to evaluate expression stability. The findings revealed that (1) the expression levels of the reference genes changed over time, even in non-differentiating cells, and (2) peptidylprolyl isomerase A (Ppia) and TATA box-binding protein (Tbp) were stable reference genes for 10 days in both undifferentiated and differentiated 3T3-L1 cells. Notably, the expression of known reference genes in non-differentiating cells was altered throughout the experiment.