title: Heart RNA-seq of control mice and a mouse model of cardiometabolic heart failure (HFpEF) treated with Rbm20 antisense oligonucleotides
design: Heart RNA-seq of control mice and a mouse model of cardiometabolic heart failure (HFpEF) treated with Rbm20 antisense oligonucleotides
library_construction_protocol: After 8 month the animals were killed by cervical dislocation and the hearts were removed, frozen in liquid nitrogen and stored at -80Â°C for further processing. The heart tissue was ground to a powder with mortar and pestle under liquid nitrogen. We used the 2-hit regimen20, in which mice were fed with a high-fat diet (D12492, Research Diet Inc) and L-NAME 0.5g/L (nitric oxide synthase inhibitor) in drinking water, beginning at 3 months of age and continue for 16 weeks. Control mice were fed with a control diet (D12450K, Research Diet Inc) and water without L-NAME. After 16 weeks on the 2-hit regimen, when the mice had fully developed the HFpEF-like phenotype, 2hit mice and control mice were randomly received Rbm20-ASOs or PBS treatment. Rbm20-ASOs were administered through subcutaneous injections at 25 mg/kg, once a week for 6 weeks. Following this treatment period (6 weeks of ASO/PBS treatment, or 22 weeks in a 2-hit regimen), these mice were employed in endpoint studies RNA from 4 left ventricles per group were isolated using TRIzol (Thermo Fisher), treated with DNAseI (Qiagen) followed by a clean-up with the Qiagen RNA micro kit.  The cDNA libraries were generated using the TruSeq Stranded mRNA Sample Prep Kit (Illumina). Both RNA integrity and cDNA library size distribution were confirmed with TapeStation (Agilent Technologies). Concentration of indexed libraries was determined using Qubit High-Sensitivity DNA kit (Invitrogen).