CTRL
Total RNA was extracted using RNeasy Mini Kit (Qiagen, Hilden, Germany). RNA concentration and RNA Integrity Number (RIN) were assessed by Agilent 2100 Bioanalyzer, RNA 6000 Nano Assay Kit (Agilent Technologies, Santa Clara, CA, USA). The cDNA libraries for RNA-seq analysis were constructed using a Sure Select Strand-Specific mRNA Library (Agilent Technologies, Santa Clara, CA, USA) according to the manufacturer's protocol. Briefly, before fragmentation, oligo d(T) beads were used to purify poly(A) mRNA from total RNA. First-strand cDNA was synthesized from the fragmented mRNA using random hexamer primers, and the cDNA libraries were prepared in accordance with the Illumina protocol. . After a purification step, the libraries were quantified with a Qubit Fluorometer (Invitrogen, Carlsbad, CA, USA) and pooled together according to their relative concentrations. The concentration and quality of each pool was assessed by Agilent 2100 Bioanalyzer. The library pools were sequenced on 4 lanes of a HighSeq 4000 (Illumina, Davis, CA, USA) with a single 1*100 bp setup using Version2 chemistry at the UC Davis Genome Center (University of California, California, USA).