The tissue was minced, homogenized, embedded in a lysing solution, and total RNA was isolated using a Macherey–Nagel kit. RNA concentrations were measured with Qubit Flourometric Quantification (Theromo Fisher Scientific), RNA integrity number (RIN) was determined with Bioanalyzer on RNA 6000 Nano Kit (Agilent Technologies), all samples were high quality with RIN > 9. RNA-seq libraries were generated using NEBNext Poly(A) + ultra–Directional RNA library Prep Kit for Illumina (New England BioLabs) according to manufacturer's instructions.