Figure 6

Characterization of different types/subtypes of spermatogenic cells by triple-staining of Ddx4, Sycp3 and Pcna. (A-J, A1-J1, A2-J2, A3-J3, A4-J4) The representative images of showing the co-staining of Ddx4 (yellow), Sycp3 (cyan), Pcna (magenta) and DAPI (grey) in the quiescent SPG-A(1-cell) (A-A4), the miotic SPG-A(1-cell) (B-B4), the SPG-A(2,4,8-cell) (C-C4), the SPG-B(16,32-cell) (D-D4), the SPG-B(64,128,256-cell), the SPC-I (L) (F-F4), the SPC-I (EZ) (G-G4), SPC-I (MZ/LZ) (H-H4), SPC-I (P) (I-I4), and the SPC-II (J-J4). Some of the indicated cells are circled by dashed lines to help identifying. (K) A schematic diagram showing the expression of the four proteins in the different types and subtypes of spermatogenic cells. SPG-A(1-cell) can be identified by the highest expression of Ddx4 and Piwil1. SPG-Bs including SPG-B(16,32-cell) and SPG-B(64,128,256-cell) are identified by the second highest expression of Ddx4 and the high expression of Pcna, while the later shows nuclear signal of Sycp3. Different subtypes of SPC-I can be identified by the expression patterns of Sycp3 and Pcna. SPD can be distinguished from SPZ by the expression of Piwil1 or Ddx4.