Single-cell RNA-seq of mus musculus: juvenile male testis
We performed testis scRNA-seq from P24 Hsf5+/+ and Hsf5-/- mice as previously described26. Briefly, testes were digested into single cell suspensions as aforementioned method for isolation of spermatogenic cells. Cell concentration was counted by Countess 3 Automated Cell Counter (Thermo fisher). Mixed the cell suspension with Trypan Blue (Thermo fisher), and then counted the viability of cells by the introverted microscope. Only samples with >80% viability were loaded into Single Cell A Chip (10X Genomics, Chromium) and generates single-cell Gel Bead-In-EMlusion (GEMs). libraries were constructed with Single Cell 3 Reagent Kits v3.1 before sequencing on the Illumina sequencing platform by Genedenovo Biotechnology Co., Ltd (Guangzhou, China).