I am using the 3T3-L1 cell line to discover the adipogenic effects of a natural compound and its derivatives. I conducted a lipid accumulation assay using fluorescent Nile Red dye (Adipored) for quantitative analysis. To support results of my lipid accumulation experiment, I also conducted gene expression analysis of adipogenic and lipogenic (A/L) genes. However, for some compounds that resulted to high lipid accumulation, they strangely have lowered or no significant change in A/L genes. On the other hand, some compounds resulted in high A/L genes but lipid accumulation were significantly lower.
Do you know any possible reason behind these results?
There are no past published papers on my treatment compounds
Hello Bryan,
Are you just measuring mRNA levels or both mRNA and protein levels? mRNA expression is not always reflective of the functional changes. It is better to measure the protein levels. Hope you will find some changes that correlate with your functional data. However, it should be kept it mind that gene expression changes are regulated at multiple levels ranging from posttranscriptional modifications, protein stability and modifications to name a few. Therefore the functional changes might be a result of other modifications of the gene rather than changes in their absolute abundance.
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