Fig 1: Differentially expressed spermatogenic proteome caused by Rpl39l deletion(A) Heatmap of differentially expressed proteins (DEPs) between wild type and Rpl39l–/– testes. Shown are three experimental repeats. DEPs are clustered into seven clusters according to their similarities in changes. Red: upregulated DEPs, Blue: downregulated DEPs.(B) Volcano plot of DEPs between wild type and Rpl39l–/– testes. Blue and red indicate top DEPs down- or up-regulated in Rpl39l–/– testes, respectively.(C and D) GO analyses of DEPs. Down-regulated GO groups relevant to ribosomes and mitochondria are shown in grey (C). Up-regulated GO groups relevant to metabolism and cytoskeleton are shown in yellow (D).(E) Western blotting of testis lysates for selected top DEPs. a-TUBULIN was used as loading control.(F) Immunostaining of testis sections. Insets show ER and acrosomal localizations of CNBP in spermatocytes and elongating spermatids from a different cross-section. Green: PNA. Blue: DAPI. Scale bar: 50 µm. Scale bar in inset: 20 µm.(G) Quantitative RT-PCR of genes encoding DEPs. Gapdh, Actin and Mrpl15 were used as controls. N = 3 testes, paired-sample Student’s t-test, *p < 0.05, **p < 0.01.(H) Changes of mRNA distributions. RNAs were extracted from sucrose gradient fractions representing RNPs, monosomes and polysomes and relative mRNAs in each fraction were compared using quantitative RT-PCR. Most mRNAs were found to accumulate in RNPs in Rpl39l–/– testes as indicated by the ratio above 1 (red dotted line). Genes encoding decreased (black) or increased (red) DEPs are indicated in G and (H)