Cellular fractionation was performed as described previously 88. Briefly, cells were washed with ice-cold PBS, collected, spun down and re-suspended in ice-cold buffer I (10 mM Hepes, pH 8.0, 1.5 mM MgCl2, 10 mM KCl, 1 mM DTT) supplemented with protease inhibitor cocktail, followed by incubation for 15 min on ice to allow cells to swell. Igepal-CA630 was then added at a final concentration of 1% (use 10% stock solution) followed by vortexing for 10 s. Nuclei were collected by centrifuging 2∼3 min at maximum speed (∼21,100 × g). The resultant supernatant was cytosolic fraction. Nuclei were then lysed in ice cold buffer II (20 mM Hepes, pH 8.0, 1.5 mM MgCl2, 25% (v/v) glycerol, 420 mM NaCl, 0.2 mM EDTA, 1 mM DTT) supplemented with protease inhibitor cocktail followed by vigorous rotation at 4 °C for 30 min and centrifugation 15 min at maximum speed. The resultant supernatant was nuclear fraction. Both cytosolic and nuclear RNAs were extracted by Phenol-Chloroform-Isoamyl Alcohol mixture (Sigma, 77618) followed by RT-qPCR analysis.

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