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.
Related Papers
- → GAPDH as a housekeeping gene: analysis of GAPDH mRNA expression in a panel of 72 human tissues(2005)841 cited
- → An old method facing a new challenge: Re-visiting housekeeping proteins as internal reference control for neuroscience research(2013)187 cited
- → Concept development of housekeeping genes in the high-throughput sequencing era.(2017)7 cited
- Relationship between the density difference of microRNA binding and the evolutionary conservation of 3'UTRs in housekeeping and non-housekeeping human genes(2013)