Single-cell transcriptomic analysis of KLF15-mediated cardiac protection in heart failure
Single-cell RNA sequencing of mouse hearts subjected to transverse aortic constriction (TAC) or sham surgery to characterize the temporal progression of cardiac remodeling from homeostasis through compensated hypertrophy to pathological heart failure. Hearts were harvested at three timepoints (5 days, 8 weeks, and 16 weeks) post-surgery to capture progressive cellular and transcriptional changes during pressure overload-induced heart failure. Overall design: Mice underwent TAC or sham surgery. Hearts were harvested at 5 days (TAC n=5, Sham n=5), 8 weeks (TAC n=3, Sham n=3), and 16 weeks (TAC n=3, Sham n=3) post-surgery (total 22 animals). Single cells were isolated via Langendorff perfusion. For the 5D and 16W timepoints, cardiomyocyte-enriched (CM) and non-cardiomyocyte-enriched (NCM) fractions were processed separately. Cells were distributed on ICELL8 250v nanowell chips and processed for scRNA-seq. Live cells were selected using Hoechst 33342 and propidium iodide staining. Libraries were prepared using the ICELL8 System and sequenced on Illumina HiSeq 4000.