Fig. 2: CEP112 Expression and Localization During Spermatogenesis and in Mature Spermatozoa. | Nature Communications

Fig. 2: CEP112 Expression and Localization During Spermatogenesis and in Mature Spermatozoa.

From: CEP112 coordinates translational regulation of essential fertility genes during spermiogenesis through phase separation in humans and mice

Fig. 2: CEP112 Expression and Localization During Spermatogenesis and in Mature Spermatozoa.

a Immunofluorescence (IF) staining of CEP112 expression across various germ cell stages in mice. (green, CEP112; blue, DAPI; Scale bar: 5 μm). b IF staining of CEP112 expression across various germ cell stages in humans. (green, CEP112; red, PNA; blue, DAPI; Scale bar: 5 μm). c Single-sperm IF analysis demonstrates CEP112 localization to the neck region of mouse spermatozoa (green, CEP112; magenta, tubulin; blue, DAPI; Scale bar: 5 μm). d Structured illumination microscopy (SIM)-based 3D reconstruction of human spermatozoa reveals CEP112 localization to atypical centrioles and the axonemal base. Lateral and axial line profiles are indicated by white dotted lines. PC, proximal centriole; DC, distal centriole. (green, CEP112; magenta, Ac-tubulin; blue, DAPI; Scale bar: 5 μm). e Immuno-gold transmission electron microscopy (IG-TEM) analysis of CEP112 subcellular localization in human spermatozoa. PC proximal centriole, DC distal centriole, Ax axoneme, Bp basal plate, Sc segmented column (Scale bar: 500 nm). f Co-immunofluorescence staining of CEP112 with centrosomal and axonemal markers (Ac-tubulin, POC1B, POC5, CEP290, or CEP63) validates CEP112 localization to atypical centrioles and axofilaments in human spermatozoa. Magnified areas are represented by dotted rectangles. Right panels, areas of co-localization represented by line-intensity profiles (green, CEP112; magenta, various markers as indicated; blue, DAPI; Scale bar: 5 μm). g Schematic representation summarizing the localization patterns of CEP112 in human spermatozoa. PC proximal centriole, DC distal centriole, Ax axoneme, Bp basal plate. h Phylogenetic analysis of CEP112 orthologs demonstrates evolutionary conservation across species. Numbers at branch points represent bootstrap values (0–100), indicating the statistical support for each branching event in the evolutionary tree of Cep112. i IF analysis of CEP112 localization in porcine and bovine spermatozoa confirms its presence in atypical centrioles. Magnified areas are indicated by dotted rectangles (green, CEP112; magenta, Ac-tubulin; blue, DAPI; Scale bar: 5 μm). Experiments were repeated three times independently with similar results; representative images are shown in a–f and i.

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