Fig. S4
Stemness test of SSCs propagated in culture. sox17::egfp SSCs that had been cultured under the propagating condition for 31 days were aggregated with dissociated testis of moto mutant, which lacks late stage germ cells. The aggregate was transplanted into rag1t26683 mutants to allow SSC maintenance and differentiation. Transplants were collected from hosts one month later for histological analysis. (A, B) Aggregates with sox17::egfp SSCs. Self-renewal of GFP-positive spermatogonia (single spermatogonium: red arrowhead; spermatogonia in small clusters: red arrow), spermatocytes (PC) and sperm (SP) were observed. (C, D) Control experiment in which dissociated moto testis was self-aggregated. Black arrowheads and arrows indicate moto-derived GFP-negative single spermatogonia or those in small clusters, respectively. (A, C) hematoxylin-eaosin staining; (B, D) GFP immunostaining. Scale bar, 20µm.