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When quantifying the mitochondrial membrane potential as a central factor of mitochondrial integrity [14], we found no impairment in the Parkin mutants under basal culturing conditions.
The biological activities mentioned above were not affected by the mutants under basal conditions (in the absence of RANTES/CCL5).
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We further determined higher levels of oxidized proteins in the mutants both under basal and stress conditions.
In contrast, the same antibody failed to detect exogenously expressed HSF1S121A mutants under both basal and metformin treatment conditions (Fig 3A).
Our results demonstrate increased oxidative stress levels in Parkin-mutant fibroblasts under basal conditions.
The expression of the gst::GFP reporter expression is increased in the cln3 mutant flies under basal conditions (Fig. 4B).
The basal level of GFP expression is ∼2-fold higher in the mutant flies, indicating that they are under an intrinsic oxidative load (Fig. 4B) and consistent with the small increase in ROS levels in mutant flies under basal conditions (Fig. 4A).
The degree of mitochondrial network branching was comparable in mutants and controls under basal conditions and decreased to a similar extent under paraquat-induced stress.
By contrast, we detected no significant differences in the degree of branching between Parkin mutants and controls under basal conditions but an increase in mitochondrial mass in the former.
First, we determined the endogenous levels of Mfn2 in the PINK1 and Parkin mutants and controls under basal conditions and after exposure to 1 µM valinomycin for 12 h.
In an earlier study, the membrane potential was found to be decreased in Parkin-mutant fibroblasts already under basal conditions and culturing in glucose depletion medium supplemented with galactose further worsened the situation [11].
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