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For Cyss depletion, cells were washed with phosphate-buffered saline solution (PBS), pH 7.4, and incubated for the indicated times in DMEM, high glucose, no glutamine, no methionine, no cystine (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% (v/v) fetal bovine serum (FBS), 0.2 mM l-Methionine, 100 g/ml streptomycin, 100 U/ml penicillin, and 2 mM l-glutamine

Maintenance Phase Period following loading where reduced dosing frequency maintains benefits without continued concentration building

The abundances of Coprobacillus , Eggerthella , and Blautia were key differentiators

GSK3 obviously increased SIRT7 phosphorylation levels in the presence of AMPK complex, whereas this effect was largely attenuated by T263A and S259A (Fig