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The level of electron transport chain uncoupling in WAT from the two genotypes was investigated by adding the chemical uncoupler FCCP.
In the execution or post-mitochondrial phase, mitochondrial membrane permeabilization results in the breakdown of the mitochondrial transmembrane potential, respiratory chain uncoupling, ATP depletion, generation of reactive oxygen species, the release of pro-apoptotic proteins into the cytosol and finally cell death.
Additionally, ROS measurements were performed using the respiratory chain uncoupling agent FCCP and the respiratory chain inhibitor rotenone.
Experiments were repeated under respiratory chain uncoupling conditions induced by 30 nmol carbonyl cyanide p- tri-fluromethoxy phenyl-hydrazone (FCCp- tri-fluromethoxy phenyl-hydrazoney chain comp- tri-fluromethoxy phenyl-hydrazone
The BAT-like phenotype of FSP27-deficient WAT was further supported by its significantly elevated expression of many genes involved in the regulation of the TCA cycle, the electron transport chain, uncoupling activity and the fatty acid oxidation pathway, resulting in its conversion from an energy storage organ to an energy consumption organ.
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Additionally, Ag+ ions could cause dysfunction of the respiratory electron transport chain by uncoupling it from oxidative phosphorylation by inhibiting respiratory chain enzymes [54].
Although UCP1 acts directly in the mitochondrial respiratory chain by uncoupling the electron transport from ATP synthesis, no apparent phenotypic alterations were observed in the P49 and P07 transgenic lines compared with the wild-type (WT) plants.
Injection of these agents enables to calculate baseline OCR, ATP turnover rate, proton leak through inner mitochondrial membrane and respiratory chain capacity (uncoupled respiration).
Our current study aims to investigate if the mitochondrial electron transport chain is uncoupled during early cancer development, and whether mitochondrial uncoupling affects p53 translocation to mitochondria.
A calmodulin-binding peptide derived from myosin light chain kinase uncouples these events.
SRC was calculated as the difference between the basal mitochondrial O2 consumption and the maximal mitochondrial O2 consumption when the respiratory chain was uncoupled by CCCP [ 30].
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