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Additional Supporting Information may be found in the online version of this article.

FilenameFormatSizeDescription
sc-12-1039_sm_SupplData.pdf12KSupplementary Data
sc-12-1039_sm_SupplFigure1.pdf363KSupplemental Figure 1.Correct timing of Blimp1-driven Cre expression in the germline. A: Representative deconvoluted fluorescence image of a whole mounted YFPRosa26loxP/ loxPBlimp1Creembryo at 7.5 dpc, merged with the corresponding phase contrast image. YFP green fluorescence is evident in visceral endoderm and in the area between the posterior proximal epiblast and the extraembryonic mesoderm where developing PGCs are found. Bar=75 μm. B: Representative paraffin-embedded section of 1 dpp testis from a LacZ -Rosa26loxP/loxPBlimp1Cre mouse after LacZ staining. Blue cytoplasmic β-Gal-positivity is evident in all gonocytes (arrows) but not in the surrounding Sertoli cell. Asteriscs point to areas of aspecific interstitial staining. Bar=30 μm
sc-12-1039_sm_SupplFigure2.pdf233KSupplemental Figure 2. Blimp1-driven Cre Sox2 deletion does not induce apoptosis within the area of PGC specification. Whole mount TUNEL assay on 7.5 dpc embryos. Upper panels show control embryos, lower panels show mutants. The arrows point to the area of PGC specification at the border between the posterior proximal epiblast and extraembryonic mesoderm. Bars=100μm
sc-12-1039_sm_SupplFigure3.pdf725KSupplemental Figure 3. Sox2 expression in PGCs and in male germ cells. Immunodetection of Sox2 in: 7.5 dpc PGCs (A-C) isolated from the posterior area of the proximal epiblast; 13.5 dpc (D-E), 1 dpp (F), 7 dpp (G) and adult (H) testis; I) shows a western blot analysis of male gonadal extracts at the indicated ages and purified postnatal germ cell extracts (sptg=spermatogonia; sptc=spermatocytes; sptd=spermatids). As a control of antibody specificity, Sox2 expression was confirmed in 13.5 dpc telencefalic ventricle (M, tv), stomach endoderm (N, en) and dorsal root ganglia (L, drg). Red and green arrowheads point to positive and negative prospermatogonia, respectively. White bars 50μm ; black bars=100μm.
sc-12-1039_sm_SupplFigure4.pdf512KSupplemental Figure 4. Blimp1-driven Sox2 deletion by Cre severely affects eye development. A-B) Bright field image of heads from control (A) and mutant (B) 1 dpp pups. (C-F) Haematoxylin eosin staining of control (C, E) and mutant (D, F) embryos sections at 17.5 dpc (C-D) or 12.5 dpc (E-F). (R=retina; l=lens). Bars=100μm.
sc-12-1039_sm_SupplFigure5.pdf585KSupplemental Figure 5. Blimp1-driven Sox2 deletion by Cre affects stomach development. A) Bright field image of control (left) and mutant (right) stomach from 1 dpp pups. B-C) Haematoxylin eosin staining of control (B) and mutant (C) stomach sections at the same age. D-M) Immunodetection of p63 (D-E) and Sox2 (F-G) and Lasp1 (H-K) in control (D, F, H, J) and mutant (E, G, I, K) stomach sections. J-K represent merged images of DAPI and anti-Lasp1 fluorescence. Bars=100μm
sc-12-1039_sm_SupplFigure6.pdf464KSupplemental Figure 6 . Blimp1-driven Sox2 deletion by Cre severely affects lung development. A-B) Haematoxylin-eosin staining of control (A) and mutant (B) lung sections from 17.5 dpc embryos. C-F) Immunodetection of Sox2 (C-D) and TTF1 (E-F) in control (C, E) and mutant (D, F) lung sections at the same age. G-H) Haematoxylin-eosin staining of control (G) and mutant (E) lung sections from 12.5 dpc embryos. Bars=100μm.
sc-12-1039_sm_SupplFigure7.pdf501KSupplemental Figure 7. Spo11-driven Sox2 deletion by Cre in meiosis does not affect germ cell development. A-B) Haematoxylin eosin staining of control (A) and mutant (B) adult ovary sections. (C-D) Haematoxylin eosin staining of control (C) and mutant (D) adult testis sections. Bars=100μm. E) Schematic representation of Floxed Sox2 locus before and after Spo11Cre mediated recombination . Arrows indicate primers used for genotyping . Non-recombined alleles do not yield amplification products. F) Genotypes of pups obtained from Sox2loxP/lacZSpo11Cre females and wt males.

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