Figure 5. Explant cultures of protrusions and posterior embryonic tissue.

Images of explant cultures. Fixed explants were stained with an antibody against Vimentin (green), Phalloidin (red) to detect Actin, and DRAQ5 (blue) to localize cell nuclei. Culture conditions supported migration and differentiation of mesoderm, including cardiac mesoderm, as evidenced by formation of rhythmically contractile centers in primitive streak explants of E7.5 embryos from normal NOD pregnancies (time-lapse video in Supplemental Information). Insets identify the area depicted in the close-up images. (a) The explant from a normal E7.5 NOD embryo had an area of rhythmically contracting myocytes (orange circle). (b) The same explant after staining, in darkfield contrast. (c) Close-up magnification of inset area. (d–f) Explant from a diabetes-exposed E8.5 NOD embryo. (g–l,m–r) Corresponding explants of posterior tissue and protrusion from the same embryo are grouped together by a green frame. Compared to posterior tissue from the same embryo, respectively, (g–i,m–o) protrusions exhibit very little outgrowth (j–l,p–r).