Mina53 antibody

ChIP assays were conducted using the EZ-ChIP^TM kit (17–371, Millipore, CA, USA) following the manufacturer’s instructions. Briefly, after sheared by sonication, the 200–1000 bps DNA fragments were immunoprecipitaed by MINA53 antibody (397300, Thermo Fisher, IL, USA) or H3K9me3 antibody (ab113754, Abcam, MA, USA), and then reverse cross-linked and purified. qPCR was conducted subsequently to determine the enrichment flod. The primer sequences used are listed in Supplementary Table S2.

Cells were lysed and proteins were extracted. Then the proteins were separated by SDS-polyacrylamide gel electrophoresis, transferred onto polyvinylidene difluoride membranes, blocked with 5% defatted milk, and incubated with the primary antibodies overnight at 4 °C followed by the secondly antibodies at for 2 h 25 °C. The ECL SuperSignal West Femto kit (34096, Thermo Fisher, IL, USA) was used for visualization. The MINA53 antibody (397300, Thermo Fisher, IL, USA), γ-H2AX antibody (#9718), p-ATR antibody (#2853), p-ATM antibody (#5883), Cleaved-Caspase 3 antibody (#9664), and Cleaved-PARP antibody (#5625) were purchased from Cell Signaling Technologies (MA, USA). The H2AX antibody (ab124781), ATR antibody (ab2905), ATM antibody (ab32420) were purchased from Abcam (MA, USA). The GAPDH antibody (AF0006) was purchased from Beyotime (Shanghai, China).