Housekeeping gene expression variability in differentiating and non-differentiating 3T3-L1 cells

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...

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Bibliographic Details
Main Authors: Danang Dwi Cahyadi, Tomoko Warita, Nanami Irie, Kana Mizoguchi, Jiro Tashiro, Yoshinao Z. Hosaka, Katsuhiko Warita
Format: Article
Language:English
Published: Taylor & Francis Group 2023-12-01
Series:Adipocyte
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Online Access:https://www.tandfonline.com/doi/10.1080/21623945.2023.2235081
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Summary: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.
ISSN:2162-3945
2162-397X