RNA-seq of mouse: adult male nasal tissue
The nasal mucosa tissues and lung tissues of AR and asthma model mice were isolated and used for RNA sequencing (n=3 in each group). RNA library preparation, sequencing, and analysis were performed by TENK Genomics (Shanghai, China). In brief, messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. The first strand cDNA was synthesized using random hexamer primers followed by the second strand cDNA synthesis, and the library was checked with Qubit and real-time PCR for quantification. After the library QC, it is pooled according to the effective concentration and the target data output required for the experiment, then subjected to Illumina sequencing, which produces 150 bp paired-end reads. Paired-end clean reads were aligned to the reference genome using Hisat2 v2.0.5, and FeatureCounts v1.5.0-p3 was used to count the reads numbers mapped to each gene.