Skeletal Muscle RNA Sequencing Protocol

Skeletal muscle RNA was obtained from all groups: vehicle, DOC, mifepristone, mifepristone plus DOC, eplerenone, and eplerenone plus DOC groups, using an EZNA^® Total RNA Kit (OMEGA Bio-Tek Inc., Norcross, GA, USA). Total RNA was quantified with a Qubit RNA BR assay kit (Invitrogen, Waltham, MA, USA). RNA quality was tested with Fragment analyzer systems (Advanced Analytical Technologies, Inc., Ames, IA, USA). Samples with an RNA quality number (RQN) greater than 9 were utilized for library construction. Then, 1 µg of RNA for each sample was used for the construction of 24 cDNA libraries using TruSeq RNA Sample Preparation kit v2 (Illumina, San Diego, CA, USA). All libraries were sequenced (2 × 150 bp) with the Hiseq X Illumina technology in Macrogen (Seoul, Republic of Korea).