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Visualization of the Epiblast and Visceral Endodermal Cells Using Fgf5-P2A-Venus BAC Transgenic Mice and Epiblast Stem Cells

Fig 2

Venus expression in Fgf5-P2A-Venus BAC Tg embryo at E6.5.

(A) Immunofluorescence analysis of the Tg embryo at E6.5 for Oct3/4 (red), Venus (anti-GFP, green), Gata4 (purple) and Nuclei (Hoechst33342, blue). Higher magnification of optical sections is shown in panels X and Y. Note that Oct3/4 and Venus were co-expressed in the epiblast of the Tg embryo, while Gata4 expression was specifically observed in the endoderm regions of the Tg embryo. Venus expression was also seen in visceral endodermal layer. Open and closed arrowheads indicate endodermal and epiblast cells, respectively. Ex: extraembryonic region; Em: embryonic region; Epi: epiblast; En: endoderm. Scale bar: 50 μm. All images were captured by a Leica TCS-SP8 confocal microscope using a 40x/1.25 oil objective lens. (B) Whole-mount fluorescence in situ hybridization of Fgf5 in the Tg embryo at E6.5. Open arrowheads indicate cytoplasmic localization of endogenous Fgf5 mRNA. Scale bar: 20 μm. Images were captured by a Leica TCS-SP8 confocal microscope using a 40x/1.25 oil objective lens.

Fig 2

doi: https://doi.org/10.1371/journal.pone.0159246.g002