title: RNA-seq of  right ventricle:Nine years female pig
design: Nuclei isolation was carried out using GEXSCOPE Nucleus Separation Solution (Singleron Biotechnologies, Nanjing, China) referring to the manufacturer's product manual. Isolated nuclei were re-suspended in PBSE to 106 nuclei per 400l, fltered through a 40m cell strainer, and counted with Trypan blue. Nuclei enriched in PBSE were stained with DAPI (1:1,000) (TermoFisher Scientifc, D1306). Nuclei were defined as DAPI-positive singlets.The concentration of single nucleus suspension was adjusted to 3-4  105 nuclei/mL in PBS. Single nucleus suspension was then loaded onto a microfluidic chip (GEXSCOPE Single Nucleus RNA-seq Kit, Singleron Biotechnologies) and snRNA-seq libraries were constructed according to the manufacturers instructions (Singleron Biotechnologies). The resulting snRNA-seq libraries were sequenced on the Illumina HiSeq X Ten system with 150-bp paired end reads.