Bulk RNA-seq of fetal and adult murine Leydig cells

To compare expression profiles of fetal and adult mouse Leydig cells, we performed bulk RNA sequencing. Overall design: Mouse testis from XY Nr5a1 Tg mouse embryos at embryonic day (E) 18.5 and 100-day old mice were harvested. Tissues were enzymatically dissociated and then filtered.The single cell solution is then sorted by fluorescent-active cell sorting to obtain a solution of alive and single Nr5a1-GFP+ cells. RNA was extracted from these cells with RNeasy Mini kit (Qiagen). Sequencing libraries were prepared with the TruSeq Stranded Total RNA Library Prep Gold (Ribo-Zero) and sequenced on an Illumina HiSeq 2500 (50 bp, paired end, ~35 million reads expected). Each condition was performed in three independent replicates.