GSM9760806: CnR,Cardiomyocytes, Malonate MI, H3K27 Rep2; Mus musculus; OTHER
For cardiomycoytes (CMs), they were obtained using the Langendorff CM isolation protocol. Cardiac fibroblasts were isolated by first digesting the hearts using the protocol from Skelly et al. () and the CFS were captured using Fluorescence-Activated Cell Sorting (FACS) with Mef-sk4 as the positive marker and Itga7 as the negative marker. Cells were then suspended in cryoppreservation buffer (10% DMSO in FBS) and stored at -80C until CUT&RUN was performed. Library construction and Cleavage Under Targets & Release Using Nuclease (CUT&RUN) assay was performed using the CUT&RUN Assay Kit (#86652S, Cell Signaling Technology) according to the manufacturer's instructions. Library quality was assessed using Agilent 2200 TapeStation with High Sensitivity D1000 ScreenTape. Sequencing libraries were prepared using the NEBNext® Ultra™ II DNA Library Prep Kit for Illumina® (NEB #E7645L) along with NEBNext® Multiplex Oligos for Illumina® (Dual Index Primers Set 1, NEB #E7600S). DNA libraries were cleaned up using AMPure XP reagent, Bechman Coulter, A63881). Libraries were sequenced on the Element Biosciences AVITI platform with 2×75 bp paired-end reads, targeting approximately 10 million reads per sample.