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The spheres were washed twice and stained with 1:500 PI solution (Bio-Sciences) supplemented with 250 g/mL RNAse A (Qiagen) and Hoechst nuclear counterstain (2 g/mL) (Bio-Sciences) for 30 min protected from light, at room temperature

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Because methylcobalamin is easier to absorb than other forms of B12, which require a conversion step prior to absorption

doi: 10.1038/nprot.2012.024