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Figure 1 | BMC Developmental Biology

Figure 1

From: A new method to transfect the hypoblast of the chick embryo reveals conservation of the regulation of an Otx2 enhancer between mouse and chick extraembryonic endoderm

Figure 1

Transfection of the hypoblast. A-E: pCAB-Luc monitored by luciferase detection at time points following hypoblast tranfection: 2 hours (A), 5 hours (B), 7 hours (C), 10 hours (D), 14 hours (E). Coloured pixels represent light intensity with blue being least intense and red most intense. F-M: GFP expression in whole embryo- (F,G) and epiblast-transfected (H,I) tissues (fluorescence images, F, H and overlaid with brightfield images, G, I). Arrowheads indicate cells expressing GFP in H. J-M: dsRed and GFP expression in the hypoblast 8 hours (J,K) and 14 hours (L,M) after transfection. An overlay of fluorescence and brightfield images are shown in K and M. N-Q: longer culture periods reveal the transfected hypoblast (shown here with GFP only) in stage HH5 (N), HH7 (O), HH8- (P) and HH9 (Q) embryos. Arrows indicate labelled cells in the yolk sac. R-S: VEcis-Otx2 driving GFP is detected in transfected hypoblasts after 7–8 hours (R) and is seen in the crescent-shaped hypoblast at stage 4+/5 (S). T-U: Co-electroporation of VEcis-Otx2-GFP and CMV-dsRed shows GFP localised in the embryo in the rostral to the hindbrain (T) and also occasionally in the hindbrain (U) (arrows). dsRed is expressed in the entire electroporated region and is seen more caudally in the embryo and in the area opaca (arrowhead).

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BMC Developmental Biology

ISSN: 1471-213X

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