Fig 9

Fig 9 Label retaining cell assays (LRC) to identify spermatogonial stem cells.

A: 32 dpf fish were treated with BrdU three times for about 16 hours per treatment (overnight) followed by a 25 day chase where fish were in water with no BrdU. At the end of the chase period, fish were fixed and processed for BrdU detection. B-F: Wild-type testes all had LRCs (B-C, arrows) with median percentage of 9.34% out of the total spermatogonia (N = 8 fish) (F). Four out of eight mutant (adad1^Y67X) testes had no LRCs (D-E) and the median percentage of LRCs of all mutant samples was less than the wild-types (F). G-I: Fluorescent in situ hybridization for nanos2 in wild-type and adad2^Y67X mutant testis. Wild-type testes had cells with bright (solid arrowhead) and faint (outlined arrowhead) nanos2 label (N = 6) (G and I). Mutant testes had either no visible nanos2 labeled cells (N = 3/6) (H and I) or had only faint nanos2 labeled cells (N = 3/6) (I). Scale bar: 20 μm.