GSM9185793: MLEC_CSE-KO_Cre-pos_H3K27ac_2; Mus musculus; OTHER
For ACT-seq library preparation, antibody–Protein A–Tn5 complexes were pre-assembled with barcoded adaptors, bound to permeabilized lung endothelial cells, and incubated to allow chromatin tagmentation. DNA was purified by phenol-chloroform extraction and ethanol precipitation, and quantified using a Qubit dsDNA HS Assay Kit (Thermo Fisher Scientific). Indexed sequencing libraries were generated by PCR amplification of tagmented DNA using Phusion High-Fidelity PCR Master Mix (NEB), with 5′ and 3′ index barcodes. Amplified products were gel-purified (QIAquick Gel Extraction Kit, Qiagen), and fragment size distribution was confirmed before paired-end sequencing on an Illumina NextSeq 2000.