Fig. 5

Loss of UHRF1 causes histone modification alteration and PIWI protein ectopic expressions. a Immunofluorescent staining with SYCP3 and H3K9me3 for surface-spread spermatocytes from Ctrl (control) and Uhrf1 cKO mouse testes. Scale bar = 5 μm. b, c ChIP-qPCR showing the H3K9me3 (b) and H3K4me3 (c) enrichments at various retrotransposons in Ctrl and Uhrf1 cKO mouse testes at P21. Quantitative data are expressed as the ratio of the ChIP (Bound) to the input DNA. Mest locus was used as a positive control for H3K9me3 and negative control for H3K4me3 enrichment. Gapdh promoter was used as a negative control for H3K9me3 and positive control for H3K4me3 enrichment. Error bars indicate the SEM of three biological replicates. *P < 0.05 by Student’s t-test. Source data are provided as a source data file. d Co-immunoprecipitation of UHRF1 followed by western blot detection of PRMT5, TDRKH, and MIWI from adult control and Uhrf1 cKO testes. e Co-immunoprecipitation of MIWI followed by western blot detection of TDRKH and UHRF1 from adult control and Uhrf1 cKO testes. f Co-immunoprecipitation of MILI followed by western blot detection of UHRF1, MIWI, and PRMT5 from adult control and Uhrf1 cKO testis extractions. g Co-immunostaining of γ-H2A.X with MILI, MIWI, TDRKH, and MVH on P18 testis sections from control and Uhrf1 cKO mice. DNA was stained with DAPI. Note: MVH (an intermitochondrial cement marker) displayed a diffused pattern in the cytoplasm of Uhrf1 cKO spermatocytes instead of perinuclear granular mitochondrial localization in controls (see zoom in on images). Scale bar = 20 μm