RNA-seq of Mst77F mutants
We collected 510 pairs of testes from 23-day-old virgin male flies and carefully separated away the accessory glands on ice. Testes were homogenized in 200 L TRIzol, followed by the addition of 40 L chloroform. After centrifugation at 13,000 xg for 15 minutes at 4 C, we collected 100 L of the aqueous phase and purified the RNA using the Zymo RNA Clean & Concentrator kit (R1015) with on-column DNase I treatment. The RNA was dissolved in 20 L ddH2O. If Nanodrop or PCR detected DNA or phenol contamination, we repeated the purification using the same Zymo kit. Purified total RNA was submitted to Novogene for poly(A) enrichment, library preparation, and sequencing on the NovaSeq X platform, generating at least 6 Gb of 150-bp paired-end reads per sample.