RNA was harvested from muscle and liver tissue using Rneasy mini kit (Qiagen) and from WAT using E.Z.N.A. Total RNA Kit II (Omega Bio-Tek). 300 ng of total RNA was used as starting material for mRNA purification and library preparation. Messenger RNA was purified from total RNA using poly-T oligo-attached magnetic beads. After fragmentation, the first strand cDNA was synthesized using random hexamer primers, followed by the second strand cDNA synthesis using either dUTP for directional library or dTTP for non-directional library. For the non-directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, amplification, and purification. For the directional library, it was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification