Fig. 4: Multi-omics analyses reveal CEP112’s role in post-transcriptional regulation of spermiogenesis-related genes.

a Gene Ontology (GO) analysis of differentially expressed proteins in Cep112^−/− testis shows significant enrichment for terms related to spermatogenesis and axonemal structures. Analysis was performed using the clusterProfiler R package (two-sided hypergeometric test; adjusted p-value < 0.05). b Gene Set Enrichment Analysis (GSEA) demonstrates significant enrichment of gene sets associated with male reproduction in Cep112^+/+ compared to Cep112^−/− testis. Statistical significance was assessed using 1000 permutations (FDR < 0.25, two-sided). c Venn diagram shows minimal overlap between downregulated genes in the transcriptome and proteome of Cep112^−/− testis. d RIP-Seq GO analysis reveals enrichment in reproductive processes (two-sided hypergeometric test; adjusted p-value < 0.05). e Motif analysis of CEP112-binding mRNAs shows specificity for known and de novo motifs. f Genome browser tracks display RIP-Seq data for Fsip2, Cfap61, and Cfap74. g RIP-qPCR confirms CEP112 binding to Fsip2, Cfap61, and Cfap74 mRNAs. Three independent experiments were performed. (Two-sided Student’s t test; error bars, mean ± SEM). The p values are indicated in the graphs. Source data are provided as a Source Data file. h RT-qPCR shows consistent mRNA levels of Fsip2, Cfap61, and Cfap74 between Cep112^+/+ and Cep112^−/− testes. n = 3 independent mice per genotype. (Two-sided Student’s t test; error bars, mean ± SEM). Source data are provided as a Source Data file. i Representative polysome gradient profiles from Cep112^+/+ and Cep112^−/− mouse testes, showing the positions of 40S, 60S subunits, 80S monosomes, and polysomes. Western blot analysis indicates CEP112 distribution across sucrose gradient fractions, predominantly in the RNP and lighter polysome regions. Three independent experiments were performed. Source data are provided as a Source Data file. j–l Polysome distribution of Cfap61, Cfap74, and Fsip2 mRNAs in Cep112^-/ testes indicates reduced translational efficiency, confirmed by quantitative analysis. (Two-sided Student’s t test; error bars, mean ± SEM). Source data are provided as a Source Data file. m TRICK reporter assay in COS7 cells demonstrates CEP112’s role in promoting CFAP61 mRNA translation. Puromycin confirms translation-dependent RFP signals. Experiments were repeated three times independently; representative images are shown.