The MMP was evaluated through JC‐1 staining, utilizing a previously established protocol.[34 (link)
] Neurons were stimulated with OxyHb and treated with or without celastrol for 24 h. After being washed 3 times with PBS, the cells were incubated in a prepared JC‐1 working solution at 37 °C for 30 min. Subsequently, the stained cells were visualized under a fluorescent microscope (Tokyo, Japan), and fluorescence intensity was quantified using a fluorescent microplate reader (excitation/emission wavelengths of 514/529 nm for monomers and 585/590 nm for aggregates) (SpectraMax Paradigm, Thermo Fisher Scientific, USA).
] Neurons were stimulated with OxyHb and treated with or without celastrol for 24 h. After being washed 3 times with PBS, the cells were incubated in a prepared JC‐1 working solution at 37 °C for 30 min. Subsequently, the stained cells were visualized under a fluorescent microscope (Tokyo, Japan), and fluorescence intensity was quantified using a fluorescent microplate reader (excitation/emission wavelengths of 514/529 nm for monomers and 585/590 nm for aggregates) (SpectraMax Paradigm, Thermo Fisher Scientific, USA).
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