Assay for transposase-accessible chromatin-sequencing (ATAC-seq) of heart samples from Wtapflox/flox and Wtap-CKO mice, respectively.

The goal of this study is to determine whether cardiac deletion of Wtap affects chromatin accessibility. To assess chromatin accessibility in the heart casued by cardiac deletion of Wtap, ATAC-seq analysis of heart samples in Wtapflox/flox and cardiomyocyte-specific Wtap knockout (Wtap-CKO) mice was performed. Each heart sample was pooled from six Wtap-CKO and seven Wtapflox/flox mice, respectively. Nuclei was extracted from heart samples, and the nuclei pellet was resuspended in the Tn5 transposase reaction mix. The transposition reaction was incubated at 37°C for 30 min. Equimolar Adapter1 and Adatper 2 were added after transposition, PCR was then performed to amplify the library. After the PCR reaction, libraries were purified with the AMPure beads and library quality was assessed with Qubit. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumina) according to the manufactuer’s instructions. After cluster generation, the library preparations were sequenced on an Illumina NovaSeq 6000 platform and 150 bp paired-end reads were generated. ATAC-seq analysis was performed using a standard protocol. The chromatin accessibility in the hearts of Wtapflox/flox and Wtap-CKO mice were characterized.