Extended Data Fig. 3: The phenotypic analysis of RibosomeST-/- mice.
From: A male germ-cell-specific ribosome controls male fertility
a, Immunofluorescence detection of LIN28 (red) with nuclei stained by Hoechst 33342 (blue), and the quantification of positively stained cells in three RibosomeST-/- strains and wild-type mice (Strain 1: P = 0.144, n = 4) (Strain 2: P = 0.759, Strain 3: P = 0.320, n = 3). Left panels scale bar, 500 μm; Right panels scale bar, 20 μm. b, Immunofluorescence detection of SYCP3 (green) and γH2AX (red) in leptotene, zygotene, pachytene and diplotene spermatocytes in RibosomeST-/- and wild-type testis and the quantification of each stage in RibosomeST-/- and wild-type mice. Scale bar, 10 μm. c, Immunofluorescence detection of SOX9 (red) with nuclei stained by Hoechst 33342 (blue), and the quantification of positively stained cells in RibosomeST-/- and wild-type mice (P = 0.161, n = 3). Left panels scale bar, 50 μm; Right panels scale bar, 20 μm. d, Fluorescence in situ hybridization of RibosomeST-/- and wild-type sperms using probes specific to X and Y chromosomes, and their corresponding ratio bar plot. Scale bar, 2 μm. e,f, Transmission electron microscopy analysis and schematic of abnormalities of RibosomeST-/- and wild-type sperms in epididymis. Red arrowhead: defects in doublet microtubules 4-7 of axoneme, blue arrowhead: other defects of axoneme. Scale bar, 1 μm. (e); The percentage of doublet microtubules 4-7 defects and other defects in axoneme of RibosomeST-/- sperm tail (P = 3.74 × 10−4, n = 3) (f). g, TUNEL staining (red) of RibosomeST-/- and wild-type testes with Hoechst 33342 stained nuclei (blue), and the quantification statistics of TUNEL positive cells per tubule (P = 0.009) and percentage of TUNEL positive tubules (P = 0.003, n = 3). Left panels scale bar, 50 μm; Right panels scale bar, 20 μm. For a, c, f and g, data are mean ± s.e.m. P values were determined using two-tailed Student’s t-tests. n values represent the number of biologically independent animals.
