The eGFP I used is the one prone to be a weak dimer. Recently, when I transfected it into HeLa cells using Lipofectamine 2000 for 24h, I noticed that it only localized in the cytoplasm (observing fixed cells). When I repeated the experiment again, I forgot to add 0.1%Triton into the PFA when fixing it. And still, the eGFP localized to the cytoplasm only. Moreover, the cells that did not express eGFP could be stained by DAPI, while the ones expressing eGFP could not. Are there any possible reasons for the phenomenon?
Does your eGFP construct have a nuclear localization signal? If fused to a nuclear protein, it could be your eGFP tag is preventing nuclear localization and therefore disrupting the function of the protein which may be involved in DAPI staining.
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