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Following 3 hours of feeding the CR mice at the start of the light cycle, all dietary groups underwent a 3 hr fast, followed by cervical dislocation and tissues for molecular analysis were flash-frozen in liquid nitrogen or fixed and prepared as described in the methods below

Similarly, the accumulation of OsVIRK1 K391E -FLAG-HA in 35S:: OsVIRK1 K391E -FLAG-HA transgenic rice plants was significantly lower than that of OsVIRK1-FLAG-HA in 35S:: OsVIRK1-FLAG-HA transgenic rice plants (Fig