Heat shock factor 5 is indispensable for pachynema progression and LINE1 silencing during spermatogenesis in mice [smallRNA-Seq]

Testes were collected from 16-day-old wild-type and Hsf5 knockout mice; small RNA libraries were constructed and sequenced on the Illumina NovaSeq 6000 platform, with adapter trimming, extraction of 18–32 nt reads for piRNA analysis, alignment to miRBase, non-coding RNAs, piRNA clusters, and transposable elements (RepeatMasker, mm39), categorization of aligned 25–32 nt piRNAs, and ping-pong analysis using LINE1 and IAP consensus sequences from Repbase, to determine whether HSF5 deficiency affects piRNA biogenesis and transposon silencing during mouse spermatogenesis by assessing piRNA abundance, length distribution, annotation profile, and ping-pong amplification in Hsf5-/- testes