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Fig. S1 nanos2 is expressed in a subset of pre-meiotic spermatogonia. (A) RNA in situ hybridization shows that nanos2 (blue staining in second row) is expressed in a subset of early germ cells, in comparison to vasa (blue staining in top row) that is expressed in all germ cells. vasa mRNA is expressed in all early spermatogonia and spermatocytes, while nanos2 expression restricted to clusters of 3–4 cells that are randomly distributed throughout the testis. (B) Fluorescent in situ hybridization for nanos2 mRNA and immunohistochemistry for Vasa protein confirms that all nanos2-expressing cells (red) are germ cells, as they are also express Vasa (green). Nuclear morphology, as revealed by DAPI staining (blue) indicates that nanos2 is expressed in a subset of pre-meiotic spermatogonia, which have characteristic uncondensed chromatin and prominent nucleoli. In A and B nanos2-expressing cells are indicated by arrows. All images are from whole mount preparations. Scale bar: 20 μm.
Reprinted from Developmental Biology, 374(2), Beer, R.L., and Draper, B.W., nanos3 maintains germline stem cells and expression of the conserved germline stem cell gene nanos2 in the zebrafish ovary, 308-318, Copyright (2013) with permission from Elsevier. Full text @ Dev. Biol.