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2.7 Enzyme-linked immunosorbent assay (ELISA) After euthanasia, mouse heart blood was collected, incubated at room temperature (23C 2C) for 2 h, and centrifuged at 13,500 g for 10 min at 4C

Vitamin D : After intestinal absorption, vitamin D3 (cholecalciferol) is transported to the liver, where it undergoes 25-hydroxylation by the enzyme CYP2R1 to form 25-hydroxyvitamin D [25(OH)D]