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Despite this probe being widely used for the measurement of mROS 36,37 , we acknowledge that there are limitations to its use, including the potential to detect changes in non-mitochondrial specific ROS 37,38

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However, studies using beta-3-adrenergic receptor knockout mice revealed a more complex picture: Long-term AOD-9604 treatment failed to produce weight loss in beta-3-AR knockout mice Acute administration still increased energy expenditure and fat oxidation in knockout mice This suggests beta-3-AR expression enhancement contributes to, but does not directly mediate, lipolytic effects The peptide appears to work through an unidentified receptor mechanism that subsequently influences beta-3-adrenergic signaling pathways