Can anyone tell me if the PGK promoter driven expression of mitoDsRed2 will be strong enough to visualise mitochondria in living cells and sort mitoDsRed2 expressing cells in FACS? Do I need a stronger promoter?
IMHO it should be just fine for sorting. Anyway DsRed2 is somewhat aggregation prone so too high expression could be detrimental. Also GFP and DsRed2 like to overlap cause they can be both excited by blue light and DsRed2 has wide spectrum and is also quite fluorescent in green wavelengths.
I worked with PGK driven expression plasmids for luciferase and gene expression and I never had an issue with too little signal or expression. Best of luck!