Transcriptomics
The mRNA was sequenced by Majorbio Bio-Pharm Technology Co. Ltd.
(Shanghai, China). In brief, total RNA was extracted from the tissue
using TRIzol® Reagent (Invitrogen, CA), followed by sample integrity,
quality, and purity accordingly. Double-stranded cDNA was synthesized
using a SuperScript double-stranded cDNA synthesis kit (Invitrogen, CA)
with random hexamer primers (Illumina). After size selection and
quantification, mRNA sequencing was performed using Illumina HiSeq
xten/NovaSeq 6000 sequencer (2 × 150 bp read length). A dataset of clean
reads aligned to reference genome was established for bioinformatic
analysis.