Fig. 6

Essential role of PRAMEF12 in spermatogonial differentiation. a Immunofluorescence of whole-mount testes from P2, P4, and P7 Pramef12^Null and Pramef12^Het mice after staining with antibodies to DDX4 (left) or KIT (middle) and merged (right). KIT-positive spermatogonia were rarely detected at P2 in either Pramef12^Null or Pramef12^Het tubules but were present at P4 and increased at P7 in Pramef12^Het tubules. Few KIT-positive spermatogonia (arrow) were observed in P7 Pramef12^Null tubules. Scale bar, 50 μm. b Immunohistochemistry of sections from P7 Pramef12^Null (top panels) and Pramef12^Het (bottom panels) testes after staining with antibodies to DDX4 (left) or KIT (middle) and merged with Hoechst 33342 to stain DNA (right). Arrowheads, KIT-positive cells in Pramef12^Null testes. Scale bar, 50 μm. c Quantitative real-time RT-PCR of Kit mRNA abundance in P7 Pramef12^Null and Pramef12^Het testes using β-actin as an internal load control and setting the abundance in Pramef12^Het testis to 1. Mean ± s.d, n = 3 biologically independent samples per condition. *P = 1.43E-05 by two-tailed Student’s t test. d Immunoblot of KIT protein in P7 Pramef12^Null and Pramef12^Het testes using α-tubulin as a load control. e Same as a, but with antibodies to SOHLH1, SOHLH2, and PLZF. The number of spermatogonia positive for SOHLH1 and SOHLH2 was significantly reduced in P7 Pramef12^Null tubules (top panels). Representative of n = 3 a, b, d, e independent biological replicates with similar results per condition