Figure 5.

(a) Quantitative RT-PCR analysis of the DEGs involved in spermatogenesis. Data represent mean ± s.d., and values were normalized to WT mRNA levels. All 21 DEGs were statistically significant (P < 0.05). (b) Western blot analysis of proteins extracted from Stra8^−/− testes at 11 dpp. Blots were probed with primary antibodies for the detection of Stra8, Rhox10, Nanos3, Egr4, Asb9, Pou5f1, Msh5 and Dmc1. The protein levels of Rhox10, Nanos3, Egr4, and Asb9 were significantly increased in HOM testes compared with WT testes (^*P < 0.05). Pou5f1, Msh5, and Dmc1 protein levels were not significantly different between WT and Stra8 knockout testes. (c) Immunohistochemistry/immunofluorescence analysis of Nanos3, Egr4, Asb9, Msh5, and Utf1 expression in WT and Stra8 knockout testes; Nanos3 (green), DAPI (blue). Scale bars = 10 μm. RT-PCR: reverse transcription polymerase chain reaction; DAPI: diamidino-phenyl-indole; s.d.: standard deviation; WT: wild-type; HOM: homozygous; dpp: days postpartum; DEGs: differentially expressed genes. Stra8: stimulated by retinoic acid gene 8; Mtl5: metallothionein-like 5; Egr4: early growth response 4; Dmc1: DNA meiotic recombinase 1; Ccdc155: coiled-coil domain containing 155; Prdm9: PR domain containing 9; Msh5: mutS homolog 5; Phlda2: pleckstrin homology-like domain, family A, member 2; Foxf1: forkhead box F1; Nkx1-1: NK1 homeobox 1; Rhox10: reproductive homeobox 10; Nanos3: nanos C2HC-type zinc finger 3; Inca1: cyclin A1 interacting protein 1; Pou5f1: POU domain, class 5, transcription factor 1; Upp1: uridine phosphorylase 1; Utf1: undifferentiated embryonic cell transcription factor 1; Asb9: ankyrin repeat and SOCS box-containing 9; Sohlh1: spermatogenesis- and oogenesis-specific basic helix-loop-helix 1; Clic6: chloride intracellular channel 6; Rrad: ras-related associated with diabetes; Mobp: myelin-associated oligodendrocytic basic protein; Pxt1: peroxisomal, testis-specific 1.