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

ABSTRACT: Normalization 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.

SUBMITTER: Cahyadi DD

PROVIDER: S-EPMC10364660 | biostudies-literature | 2023 Dec

REPOSITORIES: biostudies-literature

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

Cahyadi Danang Dwi DD Warita Tomoko T Irie Nanami N Mizoguchi Kana K Tashiro Jiro J Hosaka Yoshinao Z YZ Warita Katsuhiko K

Adipocyte 20231201 1

Normalization 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, ...[more]

PMID: 37436361

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