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dc.contributor.authorHodneland, Linn I.en_US
dc.contributor.authorPettersen, Ina Katrine Nitschkeen_US
dc.contributor.authorNikolaisen, Julieen_US
dc.contributor.authorMicklem, David Roberten_US
dc.contributor.authorDale, Hege Avsnesen_US
dc.contributor.authorRøsland, Gro Vatneen_US
dc.contributor.authorLorens, Jamesen_US
dc.contributor.authorTronstad, Karl Johanen_US
dc.PublishedScientific Reports 2015, 5:17217eng
dc.description.abstractChanges in mitochondrial amount and shape are intimately linked to maintenance of cell homeostasis via adaptation of vital functions. Here, we developed a new live-cell reporter strategy to simultaneously monitor mitochondrial biogenesis and morphology. This was achieved by making a genetic reporter construct where a master regulator of mitochondrial biogenesis, nuclear respiratory factor 1 (NRF-1), controls expression of mitochondria targeted green fluorescent protein (mitoGFP). HeLa cells with the reporter construct demonstrated inducible expression of mitoGFP upon activation of AMP-dependent protein kinase (AMPK) with AICAR. We established stable reporter cells where the mitoGFP reporter activity corresponded with mitochondrial biogenesis both in magnitude and kinetics, as confirmed by biochemical markers and confocal microscopy. Quantitative 3D image analysis confirmed accordant increase in mitochondrial biomass, in addition to filament/network promoting and protecting effects on mitochondrial morphology, after treatment with AICAR. The level of mitoGFP reversed upon removal of AICAR, in parallel with decrease in mtDNA. In summary, we here present a new GFP-based genetic reporter strategy to study mitochondrial regulation and dynamics in living cells. This combinatorial reporter concept can readily be transferred to other cell models and contexts to address specific physiological mechanisms.en_US
dc.publisherNature Publishing Groupeng
dc.rightsAttribution CC BYeng
dc.titleA new live-cell reporter strategy to simultaneously monitor mitochondrial biogenesis and morphologyen_US
dc.typePeer reviewed
dc.typeJournal article

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