Nucleoptosis: Nucleolar ISG20L2 mediates cell death through genomic DNA degradation

In this study, we employed ChIP-seq to identify ISG20L2 directly interacts with genomic DNA during nucleoptosis in A549 cells A549 cells were infected with HSV-1 (MOI: 1) for 8 hours, and then collected, fixed and undergone chromatin cross-linking according to the manufacturer’s instructions using the SimpleChIP® Enzymatic Chromatin IP Kit (9003S, CST). The chromatin fragments specifically binding to ISG20L2 were enriched by anti-ISG20L2 antibody (24639-1-AP, Proteintech), and then the DNA was purified by QIAquick PCR Purification Kit (28104, QIANGEN). The purified DNA was then constructed for ChIP-seq library by Novogene (Beijing, China). The clean reads were aligned to human genome (GRCh38.p14) using BWA mem (v 0.7.12), and the IP enriched peaks were then analyzed by MACS2 (version 2.1.0).