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Overexpression of MIC26 induced fragmentation of mitochondria, promoted ROS formation and resulted in impaired mitochondrial respiration.
Importantly, we found that Trak1 depletion also caused fragmentation of mitochondria into small tubules and spheres (Fig. 1B and 1C), indicating that endogenous Trak1 is required for normal morphogenesis of mitochondria.
The change in fission/fusion balance impacts mitochondrial function, and LPS has been shown to affect this dynamics by upregulating the fission protein dynamin-related protein 1 (Drp1), which results in disrupted distribution and fragmentation of mitochondria [32].
TMRE staining intensity was similar in control and PHB1-depleted cells (Fig. S2A), suggesting that the mitochondrial membrane remains intact despite fragmentation of mitochondria in these cells.
After 5 days, cells from the central region (thus older cells) showed a drastically increased fragmentation of mitochondria and more punctured structures whereas freshly nourished younger cells from the outer rim displayed a normal tubular structure of mitochondria (Fig 1E).
Thus, monitoring of cell death by confocal microscopy revealed the following events: intracellular ROS generation concomitant with fragmentation of mitochondria, ΔΨm drop and subsequent loss of plasma membrane permeability for PI.
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Here, we first investigated the effect of C60 COOH on mitochondria dynamics, showing that C60 COOH treatment prevents mitochondria fragmentation, depolarization of mitochondria membrane potential, as well as attenuation of NF-κB and MAPK activation, thus leading to reduced neuroinflammatory response.
Now we show that our approach can also be applied to improve the estimation in the fragmentation level of mitochondria in a single cell without actually identifying each mitochondrion object in a cell image.
Specifically, calcineurin-dependent dephosphorylation of Ser637 effects Drp1 translocation to mitochondria, creating a loop in which fragmentation of depolarized mitochondria involves intracellular Ca2+ increase, activation of calcineurin, and dephosphorylation and translocation of Drp1.
In apoptotic cells, the fragmentation of depolarized mitochondria depends on Ca2+-evoked, calcineurin-mediated dephosphorylation of Drp1 at its conserved serine 637 site (Cereghetti et al., 2008).
In apoptotic cells, the Ca2+-evoked activation of calcineurin mediates the dephosphorylation of Drp1 at its conserved serine 637 site, triggers the translocation of Drp1, and results in the fragmentation of depolarized mitochondria (Cereghetti et al., 2008).
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