GSM8182838: HpNSCs, IgG, CUT&Tag; Mus musculus; OTHER
CUT&Tag was performed using the NovoNGS CUT&Tag 3.0 High-Sensitivity kit (N259-YH01, Novoprotein) according to the manufacturer's instructions. Briefly, 1x105 HpNSCs were harvested, immobilized and permeabilized with NovoNGS ConA Beads. Cells were incubated with GR antibody (Santa Cruz, sc-1004) or IgG control antibody (Millipore, 12-370) overnight at 4 °C, followed by incubation with an unlabeled secondary antibody (Millipore, AP132) for 1 h at room temperature. Subsequently, pAG-Tn5 was added to the samples for 1 h. The cells were then incubated in Tagmentation Buffer at 37 °C for 1 h. DNA fragments were purified with the Tagment DNA Extract Beads. Libraries were amplified by PCR following the provided cycling conditions by the NovoNGS CUT&Tag 3.0 High-Sensitivity kit. Library quality was assessed using an Agilent Bioanalyzer, and sequencing was performed on an Illumina Novaseq 6000 platform.