title: GDF15 antagonism limits severe heart failure and prevents cardiac cachexia in mice
design: GDF15 antagonism limits severe heart failure and prevents cardiac cachexia in mice
library_construction_protocol: The fresh hearts were harvested and snap frozen in liquid nitrogen.Meanwhile for 7. mice, tissues were harvested and immediately snap frozen in liquid nitrogen and stored at _80_C until further analysis. Animals were perfused with cold PBS to remove any trace of blood. Aortas were harvested and cleaned of contaminated tissues under a dissection microscope mouse 6-8 weeks Pppr15a-/- with irradiation Align to GRCm39 reference genome using star (version 2.6.1d) RNA was then extracted (and in some instances treated with DNase1 on-column) using the RNeasy isolation kit following the manufacturerâ€™s instructions. For RNA isolation, approximately 30-50 mg of tissue was placed in Lysing Matrix D tubes and homogenized in 800 ml Triazol (Qiazol,QIAGEN) using the Fastprep-24 Homogenizer for 30 s at 4-6 m/s (MP Biomedical), or a rotor-stator homogenizer. The resultant supernatant was transferred to an RNase free tube and 200 ml chloroform (Sigma) added. The samples were vortexed and centrifuged at 13 000 rpm for 15 min at 4_C. The upper phase was then transferred to a RNase free tube and mixed with equal volume of 70% ethanol before loading onto RNA isolation spin columns (QIAGEN).