Fig. 6

The synergistic enhancement between Pramex1 and Pramel1 was through retinoic acid (RA) signaling pathway. A Representative images of whole-mount IFS with TRA98 (red) on seminiferous tubules of WT, Pramex1 sKO, Pramel1 sKO, and Pramex1/Pramel1 dKO mice at P7, these animals were treated with RA or WIN18,446 (RA inhibitor) at P2. TRA98 was used to label germ cells. The SCO regions, labeled with white dashed line, lacked germ cells which were seminiferous tubules without any TRA98 + cells. Nuclei counterstained with DAPI (blue). Bar = 100 µm. B SCO segments along the length of seminiferous tubules (%) based on the whole-mount IFS for the RA/WIN18,446 treatment in the four groups (n = 3). Significance was assessed among the four groups (WT, Pramex1 sKO, Pramel1 sKO, and Pramex1/Pramel1 dKO mice) for each treatment (RA or RA inhibitor). C Co-immunoprecipitation (co-IP) analyses of PRAMEX1 and RARα in P7 WT, Pramex1 sKO and Pramel1 sKO testis tissue. YBX2 served as a control. D Based on the band intensity in the WB for WT testis tissue, the protein expression level relative to input WT testis (set as 1) was measured (n = 3). Significance was assessed among the three samples (input, IP/w PRAMEX1, and IP/w RARα) for each antibody (PRAMEX1, RARα and YBX2). E Based on the band intensity in the WB, the protein expression level relative to input Pramel1 sKO testis (set as 1) was measured (n = 3). Significance was assessed among the three samples (input, IP/w PRAMEX1, and IP/w RARα) for each antibody (PRAMEX1, RARα and YBX2). Data were expressed as mean ± SEM. Values that do not share a common superscript (a–d) were found to differ significantly (P < 0.05)