Rat caput epididymis scRNA-seq
Cell suspension preparation was conducted as previously reported. In brief, tissues were washed three times using Hanks balanced salt solution and minced into pieces. The tissues were digested, and then a 4 m sterile strainer (Corning Incorporated, Tewksbury, MA, US) was used to remove cell debris and other impurities. The cells were collected by centrifugation at 500 g at 4 C for 5 min. After that, the cells were resuspended in PBS and counted with a TC20 automated cell counter (Bio-Rad). Cellular suspensions were loaded on a 10  Genomics GemCode single-cell instrument that generates single-cell gel beads-In-EMlusion (GEMs). The gel beads in a GEM were dissolved to release barcode sequences, and the barcoded full-length cDNAs were reverse transcribed and amplified by PCR to generate sufficient mass for library construction. The purified scRNA-seq libraries were sequenced on an Illumina HiSeq X Ten system with 150 bp paired end reads.