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Cell cytoplasmic/nuclear fractionation
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Cytoplasmic and nuclear fractions were isolated via mild lysis and centrifugation using the Nuclear/Cytosol Fractionation Kit (Biovision, Milpitas, CA). Briefly, 2 × 106 cells were treated with trypsin-EDTA (Gibco) and resuspended with cytosol extraction buffer A. Cytosol extraction buffer B was added to the suspension followed by centrifugation to obtain cytosolic protein/RNA containing supernatant, whereas the pellet was further processed for nuclear proteins/RNAs.
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