Knockdown of KIT⁺-enriched KZFPs results in derepression of ERVs and their adjacent genes. (A,E) Schematic of ERV enhancers and their target loci. (B,F) Results of qRT-PCR for KZFP gene-targeted ERV and ERV-adjacent genes under Zfp987 knockdown (B) and Zfp989 knockdown (F). Data values for controls were normalized to 1.0. Student's t-test. (**) P < 0.01, (*) P < 0.05. n = 4. (C,D,G) Track views of KZFP, RNA-seq, H3K9me3, and H3K27ac enrichment in KIT⁺ and PSs at the target ERV loci. Genomic regions around target genes (Or1f19 [C], Vmn2r100 [D], and Ube3a [G]) are shown. Target ERVs are represented as red-colored letters and quadrangles. Adjacent regions of ZFP987 or ZFP989 binding sites are highlighted with gray-colored areas. (H) Summary: In spermatogonia, spermatogonia-enriched KZFPs bind and suppress ERVs that later function as enhancers. At the ERV loci targeted by spermatogonia-enriched KZFPs, after entering meiosis, KZFPs were displaced and H3K9me3 decreased, whereas H3K27ac increased to become active enhancers.