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Tissue-Specific Stem Cells
Version of Record online: 5 JUL 2013
Copyright © 2013 AlphaMed Press
Volume 31, Issue 7, pages 1408–1421, July 2013
How to Cite
Campolo, F., Gori, M., Favaro, R., Nicolis, S., Pellegrini, M., Botti, F., Rossi, P., Jannini, E. A. and Dolci, S. (2013), Essential Role of Sox2 for the Establishment and Maintenance of the Germ Cell Line. STEM CELLS, 31: 1408–1421. doi: 10.1002/stem.1392
Author contributions: S.D.: conception and design; S.D., E.A.J., and P.R.: financial support; M.P.: administrative support; S.N., R.F, and M.P.: provision of study material or patients; F.C., M.G., F.B., E.A.J., and S.D.: collection and/or assembly of data; F.C., M.G., P.R., and S.D.: data analysis and interpretation; S.D. and P.R.: manuscript writing; S.D.: final approval of manuscript. F.C. and M.G. contributed equally to this article.
Disclosure of potential conflicts of interest is found at the end of this article.
first published online in STEM CELLS EXPRESS April 3, 2013.
- Issue online: 5 JUL 2013
- Version of Record online: 5 JUL 2013
- Accepted manuscript online: 3 APR 2013 05:23AM EST
- Manuscript Accepted: 13 FEB 2013
- Manuscript Revised: 28 JAN 2013
- Manuscript Received: 5 NOV 2012
- Italian Ministry of University. Grant Numbers: Prin 20084XRSBS_004, Prin 2009FW5SP3_001, Firb RBAP109BLT_004
- Cariplo Foundation. Grant Number: 20100673
- ASTIL Regione Lombardia. Grant Numbers: SAL-19, N Prot FL 16874
- Telethon. Grant Number: GGP12152
- Associazione Italiana Ricerca sul Cancro. Grant Number: AIRC IG-5801
Additional Supporting Information may be found in the online version of this article.
|sc-12-1039_sm_SupplFigure1.pdf||363K||Supplemental 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.pdf||233K||Supplemental 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.pdf||725K||Supplemental 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.pdf||512K||Supplemental 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.pdf||585K||Supplemental 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.pdf||464K||Supplemental 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.pdf||501K||Supplemental 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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