Supplementary Information for
Hu YC, Nicholls PK, Soh YQ, Daniele JR, Junker JP, van Oudenaarden A, Page DC.
Licensing of primordial germ cells for gametogenesis depends on genital ridge signaling
PLOS Genetics 11, e1005019 (2015)
- Figure S1 Post-migratory, but not migratory, germ cells expressDazl. (A) Immunofluorescentstaining for SSEA1 and smFISH forDazlandOct4at E9.5.Dazlexpression was not detectable inmigratory PGCs, whileOct4was. Individual germ cells are outlined by dashed lines. (B) Represen-tative E11.5 whole-gonad image ofDazlandOct4transcript molecules marked by smFISH.Dazltranscript counts are higher in anterior region than in posterior region.Dazl(pseudocolored inblue, upper panel) andOct4(pseudocolored in red, lower panel) mRNA molecules were overlaidwith DAPI staining (white) and identified after raw images were subject to image analysis filtering.Raw images prior to filtering are displayed in magnified images (middle panels), where mRNAmolecules are visible as white dots within individual germ cells (outlined by dashed lines).
- Figure S2 Germ cells inGata4cKO (soma-specific Cre) embryos do not express DAZL orMVH. Immunofluorescent staining for SSEA1, DAZL, MVH, and GATA4 in transverse sections of control andGata4cKO (Osr1CreER) embryos on a C57BL/6 genetic background.Nuclei counterstained with DAPI (blue). Scale bars: 50Ám.
- Figure S3 Germ cells inWt1-deficient genital ridges express DAZL. Genital ridge formation isinitiated inWt1KO embryos (a gift from Kenneth H. Albrecht), but growth is severely retardedand degeneration ensues. Immunofluorescent staining of longitudinal sections from wildtypeorWt1KO urogenital regions shows that PGCs at the genital ridge (GATA4-positive, blue) ex-press DAZL (red, arrows). Yellow dashed lines outline the genital ridge. Autofluorescent redblood cells are indicated (asterisk). gr, genital ridge. Scale bars: 50Ám.
- Figure S4 Germ cells inOsr1-deficient genital ridges express DAZL. Genital ridge formation isinitiated inOsr1KO embryos, but complete degeneration occurs by E15.5 . (A) Immuno-histochemical staining for GATA4 in cross-sections of wildtype andOsr1KO embryos atE11.5. Genital ridge formation is initiated inOsr1KO embryos, but growth is restricted. Insetshows higher magnification of genital ridge. (B) Immunofluorescent staining for SSEA1,DAZL, and GATA4 in cross-sections of wildtype andOsr1KO urogenital regions. Representa-tive germ cells positive for DAZL are indicated by arrows. Yellow dashed lines outline the geni-tal ridge. a, dorsal aorta; gr, genital ridge; m, mesentery. Scale bars: 50Ám.
- Figure S5 Germ cells inGata4cKO (soma-specific Cre) cultured UGRs do not express DAZL or MVH. Immunofluorescent staining for SSEA1, DAZL, MVH, and 5-methyl-cytosine (meC)in transverse sections of control andGata4cKO (Wt1CreER;Osr1CreER) cultured UGRs (on aC57BL/6 genetic background). Nuclei counterstained with DAPI (blue). Inset shows highermagnification of germ cells. Scale bars: 50Ám.