I've transfected fibroblasts with GFP tagging mitochondria and I collected the extracellular vesicles secreted . usually with my non transfcted cells, I used to see mitochondrial material in my EVs so I was expecting getting extracellular vesicles tagged with GFP but it was not the case for my EVs but I did see GFP in my transfected cells, how do you think can I get fluorescence in my EVs?
Hi Amari,
If we are in the same, you're planning to visualize GFP on your EVs since they can contain GFP tagged mitocondria. But It didn't happen, I can suggest you to use a GFP with a signal peptide to localize on/in endoplasmic reticulum. An easy tip is avoid extra washing steps to not get rid of GFP-tagged mitochondria.
What signal Peptide did you use for GFP ?
Have you used a higher concentration of GFP plasmid for transfection and check for GFP tagged mitochondria in the EVs? This is again at concentrations that do not alter mitochondrial or cellular health. You may try (if not done yet) and revert on whether it worked.
Noe baruch@ i used plasmid with a presequence tagged for soluble matrix mitochondrial proteins
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