GSM8266896: Adult CF, shNr4a3_1, H3K27me3 CUT&Tag; Mus musculus; OTHER
50000 cells were harvested, and the CUT&Tag assay was performed by SeqHealth (Wuhan Seqhealth Co., Ltd. China) using a commercial kit (TD903, Vazyme, China) according to the manufacturer's protocol. The following primary antibodies were used: Rabbit anti-H3K27ac antibody (Abcam; ab4729), anti-H3K27me3 antibody (Active Motif; Catalog No: 39155). Hyperactive pG-Tn5 transposonase was then added to perform tagmentation. The reaction was terminated, and DNA fragments were extracted by PCI, followed by PCR amplification using indexed P5 and P7 primers. cells were incubated with Rabbit anti-H3K27ac antibody (Abcam; ab4729) and anti-H3K27me3 antibody (Active Motif; Catalog No: 39155) for 2 h, and then 1 h with secondary antibody at room temperature. Hyperactive pG-Tn5 transposonase was then added to perform tagmentation. The reaction was terminated, and DNA fragments were extracted by PCI, followed by PCR amplification using indexed P5 and P7 primers. The library products were enriched, quantified and finally sequenced on a Novaseq 6000 sequencer (Illumina) with PE150 model.