aNSCs differentiation assay After 24 h of incubation in the proliferation medium, half of the medium was replaced with differentiation medium containing Neural Stem Cell Basal Medium, B27 without vitamin A, GlutaMAX, antibiotics (100 IU/mL penicillin and 100 g/mL streptomycin), 1 mg/mL epidermal growth factor, 1 mg/mL fibroblast growth factor 2, 1 M retinoic acid (R2625, Sigma-Aldrich), and 1 M forskolin (F6886, Sigma-Aldrich)
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Add 8 ml 0.05 molL -1 sodium phosphate buffer (pH = 7.8) precooled in advance, centrifuge at 10000 rpm for 20 min, and store the supernatant at 4C for enzyme activity determination ( 2.4 Untargeted metabolomics of buckwheat leaf tissues Untargeted metabolomics, a useful approach for the simultaneous analysis of samples with significant phenotypic changes, has been used to detect metabolites after Cd treatment ( Non-targeted metabolic profiling analyses were profiled by Perkin Elmer 680 GC (Perkin Elmer Inc, Akron, OH, USA) and Q Exactive Focus Orbitrap LC-MS/MS (Thermo Scientific, Waltham, MA, USA)