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

FilenameFormatSizeDescription
DVDY_22761_sm_suppinfo.pdf26KSupporting Information
DVDY_22761_sm_suppinfofig1.tif179KSupporting Information Fig. S1. B1-RESC sex determination. To determine B1-RESC sex real-time, PCR from retro-transcribed mRNA was performed. Primer sequences were designed in the sex-determining region Y (Sry) (Supp. Table S5). As controls, cDNA samples from known male and female rat tissues were run in parallel. PCR products run in a 2.5% agarosa gel are included. A DNA ladder of 100-bp fragments was used.
DVDY_22761_sm_suppinfofig2.tif82KSupporting Information Fig. S2. Thyroid hormone alpha-1 receptor expression. B1-RESCs express Thyroid hormone alpha-1 receptor (TRα1) as studied by immunocytochemistry in single cells from EBs after 2 days in vitro in spontaneous differentiation conditions (no MEF, no mitogens). Magnification, 60×. EBs, embryoid bodies.
DVDY_22761_sm_suppinfofig3.tif154KSupporting Information Fig. S3. p75 low-affinity NGF receptor expression. B1-RESC single cells from EBs after 2 days in vitro (spontaneous differentiation) show p75 inmunoreactivity. Magnification 60×. The expression of p75 was also studied by real-time PCR. EBs, embryoid bodies.
DVDY_22761_sm_suppinfofig4.tif7403KSupporting Information Fig. S4. B1-RESC Endoderm and Mesoderm markers expression. The expression of laminin, alfa fetoprotein (representative markers for endoderm), and BMP4 (representative marker for mesoderm) was studied by inmunocytochemistry in single cells obtained from EBs. Representative images of B1-RES single cells immunoreactive for laminin (A), alfa fetoprotein (B), BMP4 (C), and vimentin (D) at 10 days in vitro. Hoechst 33258 nuclear staining was also performed. Magnification, 60×.
DVDY_22761_sm_suppinfofig5.tif106KSupporting Information Fig. S5. B1-RESC ectoderm, endoderm and mesoderm markers expression. The expression of markers representative of the three germ layers was also studied by real-time PCR in single cells obtained from EBs after growing in spontaneous differentiation conditions. Amplified PCR products after using appropriate primers for nestin (ectoderm), Sox-17 and FOXA2 (endoderm), and DKK1 (mesoderm) are included. GAPDH product of PCR in the similar cDNA samples has been also included. No template controls (NTC) and no RT samples were run in parallel to PCR reactions.
DVDY_22761_sm_suppinfotab1and2.pdf16KSupporting Information Tables 1 and 2.
DVDY_22761_sm_suppinfotab3and4.pdf25KSupporting Information Tables 3 and 4.
DVDY_22761_sm_suppinfotab5.pdf12KSupporting Information Table 5.

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