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...
Main Authors: | , , , , , , |
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Format: | Article |
Language: | English |
Published: |
Taylor & Francis Group
2023-12-01
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Series: | Adipocyte |
Subjects: | |
Online Access: | https://www.tandfonline.com/doi/10.1080/21623945.2023.2235081 |
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. |
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ISSN: | 2162-3945 2162-397X |