GSM3664687: A77 1726 25uM_r3; Homo sapiens; OTHER
Cells were washed in PBS. Cells were detached using accutase and collected in 50ml tube. Cells were spin 300gx4mins washed with washing buffer :10mM Tris-Cl, pH 8.0, 10mM KCl, 250mM sucrose, 5mM MgCl2, 0.5mM DTT, 10% Glycerol. Cells were spin again and then resuspended in permeablization buffer (Washing+0.1% Igepal). Cells were incubated 2mins at RT and then spin again and resupended in Freezing buffer (50mM Tris-CL pH 8.3, 40% glycerol, 5mM MgCl2, 0.5mM DTT. Nuclei were counted and aliquoted in 1million/tube and flash frozen. using 1 million cells per sample. Samples were spiked with 40,000 Drosophila S2 cells as a normalization spike-in control. Samples were sequenced on the NextSeq using a High Output 75-cycle kit, generating paired end 42 nt reads. Libraries were sequenced to an average read depth of 10 million mappable reads per sample. Pro-Seq