BRD4 inhibitor suppresses melanoma metastasis via the SPINK6/EphA2-EGFR pathway

Chromatin immunoprecipitation (ChIP)-seq was performed by Acegen. Briefly, A375 cells were treated with 4 nM NHWD-870 for 3 days or transfected with BRD4 siRNA. Then, those cells were collected and cross-linked in DMEM with 1% formaldehyde. A bioruptor was used to sonicate chromatin, followed by incubation with BRD4 antibody (#13440, CST) for immunoprecipitation. The DNA library was prepared using the Acegen DNA Library Prep Kit from Illumina, then twelve-cycle PCR amplified, cleaned up, analyzed by an Agilent 2100 Bioanalyzer and finally sequenced on the Illumina platform.