title: RNA-Seq of lungs: Juvenile male and female sheep
design: Total RNA was extracted using TRIzol reagent (Thermo Fisher) according to the manufacturer's instructions. The concentration and integrity of isolated RNA were assessed using a NanoDrop 2000 UV (Thermo Fisher) and 1% agarose gel electrophoresis, with 18 S and 28 S ribosomal RNA serving as a reference. High-quality RNA samples were sent to Sangon Biotechnology Co., Ltd. for cDNA library preparation and RNA sequencing on the Illumina HiSeq platform. After sequencing, raw reads with adapter, and poly-N and low-quality reads (QV < 20) were discarded. Clean reads with high quality were used for de novo transcriptome assembly using Trinity software. Gene Ontology (GO), NCBI nonredundant protein sequences (NR), Protein family (PFAM) and euKaryotic Ortholog Groups (KOG), and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used for transcript annotation. All transcript abundances in this study are represented using transcripts per million (TPM) values, and TPM with q-value <0.05 and |FoldChange| >2 among groups were used as cut-off criteria for differentially expressed genes (DEGs) using the DESeq2 R package.