Total RNA was extracted from frozen lung tissues using Trizol reagent (Invitrogen), solubilized in UltraPure RNase/DNase-free water, and quantified by using NanoDrop ND-1000 spectrophotometer (Marshall Scientific, Hampton, NH). For RNA-seq analyses, 1 µg total RNA was used as starting material. First,ribosomal RNAs were depleted using a NEBNext rRNA Depletion Kit (New England Biolabs, Ipswich, MA), according to the manufacture's recommendations. The RNA library was prepared using a NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs). Briefly, RNA was fragmented, and cDNA was synthesized using random primers. The resulting double-strand cDNA was then subject to end-repair, adapter ligation, and PCR amplification to generate each library.
GEO Accession
GSM3777071