Fig. 1

Pramef12 is essential for spermatogenesis and male fertility. a The hierarchy of different cell types of spermatogonia during SSC self-renewal and differentiation. The A_s spermatogonia are heterogeneous with SSCs and A_s progenitors. The A_s and A_pr progenitors have the potential to become SSCs. A_s spermatogonia produce chains of A_pr A_al(4), A_al(8), and A_al(16) undifferentiated spermatogonia that are connected by cytoplasmic bridges and are precursors of differentiated spermatogonia. b Fertility of > 3 pairs of Pramef12^Null and Pramef12^Het male and female mice mated 1:1. Mean litter sizes ± s.d. are shown with indicated genotypes. c Testes of P90 adult Pramef12^Null and Pramef12^Het mice. Scale bar, 1 mm. d Ratios of testis to body weight of Pramef12^Null and Pramef12^Het mice shown in c. Mean ± s.d, n = 6 biologically independent testes from six different animals. *P = 1.44E-08 by two-tailed Student’s t test. e Adult testis sections from Pramef12^Null and Pramef12^Het mice stained with periodic acid-Schiff (PAS) and hematoxylin. Pramef12^Null testes are agametic with a Sertoli cell-only phenotype. Scale bar, 50 μm. f Immunofluorescence of P90 adult testes from Pramef12^Null and Pramef12^Het mice after co-staining with antibodies to DDX4 (germ cells) and WT1 (Sertoli cells) as well as Hoechst 33342 (DNA). Scale bar, 50 μm. g Immunohistochemistry of P90 adult testes from Pramef12^Null and Pramef12^Het mice after staining with antibodies to cyclin D1 and PLZF. Arrowheads indicate positive-staining spermatogonia. Scale bar, 50 μm. h Quantification of cyclin D1-positive and PLZF-positive spermatogonia in Pramef12^Null and Pramef12^Het testes. Mean ± s.d, n = 3 biologically independent testes from three different animals. *P = 1.44E-28 (cyclin D1) and *P = 3.19E-26 (PLZF) by two-tailed Student’s t test. i Same as e, but of cauda epididymides. Representative of n = 6 c, n = 3 e, f, g, i independent biological replicates with similar results per condition