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. 2009 Oct 28;4(10):e7650. doi: 10.1371/journal.pone.0007650

Figure 3. Two hours of contact between animal cap and anterior endoderm is sufficient for cardiogenesis.

Figure 3

(A) Conjugates were made between st. 10.25 anterior endoderm explants and animal caps of aged animal caps isolated at st. 8.5, and MHCα expression was analyzed a day and a half later, at st. 34. (B) Conjugates were made between st. 8.5 animal caps and aged anterior endoderm explants isolated at st. 10, as indicated. MHCα expression was analyzed at st. 34. (C) Strategy for a peeled animal cap assay. Rhodamine-dextran injected anterior endoderm is conjugated with animal caps, and the caps are removed after 1 or 2 hours. Anterior endoderm-free (rhodamine-negative) animal caps are cultured until st. 34 or for a total of 2.5 hours after the initial contact, corresponding to st. 10.5 (D) Only animal caps that were in contact with anterior endoderm for 2 hours express MHCα, but other markers tested – Msr, Globin, Edd – are also induced after 1 hour of contact. (E) Early markers of endoderm (Sox17, Hex, Cerberus, Mixer), Organizer (Goosecoid (Gsc), Chordin) and mesoderm (Xbra, Eomesodermin) are expressed after 1 hour contact with anterior endoderm. However Mesp1, a marker of migrating cardiac precursors [62], was not induced. (F) Myogenic regulatory factors MyoD and Myf5 as well as posterior markers Xpo and Vent2 are not induced by AE.