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Separation of soluble and insoluble components Single mouse hippocampal tissue was isolated, and 300 l mixed protein lysis solution with RIPA (Applygen, C1053), a protease inhibitor (Merck, 539134), and a phosphatase inhibitor (Merck, 524628) (=100:1:2) were added

A quantity equivalent to 60 mg of l-glutathione was weighed and transferred in to 10-mL volumetric flask

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Subsequent flow cytometric analysis of NRF2 expression in ILC2s revealed that melatonin treatment significantly increased NRF2 protein levels ( Figure 6D )