title: GSM6974770: CM_control_2; Mus musculus; RNA-Seq
library_construction_protocol: MX1gfp mice injected with vehicle or Doxorubicin were anaesthetized using isoflurane vaporizing system, and anesthesia was maintained using a nose cone throughout the surgical procedure. Following anesthesia, thoracic cavities were opened to expose the heart. Descending aorta was severed to allow exsanguination, and 7 mL EDTA buffer was immediately injected into the right ventricle. Ascending aorta was clamped using a hemostat, and the heart was removed from the mice then transferred to a 60 mm dish containing EDTA buffer. The heart was again injected  with 10 mL EDTA buffer into the left ventricle at 2 to 3 mm above apex. Next, the heart was transferred to a 60 mm dish containing perfusion buffer and injected with 3 mL of perfusion buffer through the same perforation on the left ventricle. Lastly, the heart was transferred to a 60 mm dish containing collagenase buffer, and heart digestion was achieved by continuously injecting collagenase buffer through the same perforation at the flow rate approximately 2 mL/min until the heart turned pale and flaccid and the injected perforation was enlarged. The atria and ventricles were gently separated and then the ventricles (with septum) were transferred to a new 60 mm dish containing 3 mL fresh collagenase buffer. The digested tissues were pulled into 1 mm pieces using fine-tip forceps, and cardiac cells were dissociated by gentle trituration, and then the enzyme activity was neutralized by adding 5 mL stop buffer. The single cell suspension was filtered through a 70 µm strainer, and the myocytes were settled by gravity for 20 min in a 50 mL tube. Direct-zol RNA miniprep kit (Zymo Research) was used to total RNA for sequencing. RNA libraries were prepared for sequencing using standard Illumina protocols