Great.. never thought about it. But can you please tell me why such later time point can help to answer or cite any publication over it?
We have seen a similar effect with some mRNAs//proteins. I think the idea that the half -life of the protein may explain this is an interesting suggestion.
If you have an experimental model where over-expression (or under-expression) of an miRNA is stable for the ensuing 24hrs or more this possibility could be investigated.
yes I have done luciferase assay followed by qPCR and WB. Luciferase and qPCR showed decrease of target expression whereas WB does not. I thought luciferase assay results correlate with WB but thats not my case. any clue whats happening?
We have examined the RNA transcriptomes following transfection with one single miRNA mimic and found that 20-30 other miRNAs, as well as many other mRNAs, exhibited significantly altered expression in the transfected cells (versus scrambled control). Considering the multiplicity of miRNA targeting it therefore possible, even likely, that the 1:1 inverse effect will not always be observed.
Besides the time course (for miR, mRNA and protein) I would check the other putative targets for your miR and how they interact with your addressed protein. A signaling loop could counteract the negative effect you expect. miR usually behave pleiotropically and more one gene are affected. But, the most likely reason is the time-dependent effect.
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