Figure 3

Methylated DNA nuclear patterns at early in vitro embryogenesis of microspore and immature zygotic embryo. A-D: Microspore embryogenesis. E-I: Somatic embryogenesis of immature zygotic embryos. A, A’, E, E’: Confocal images of 5mdC immunofluorescence in early multicellular embryos (emb) and embryogenic masses (ms). Differential interference contrast, DIC, images (A, E), and merged images of 5mdC immunofluorescence (green) and DAPI staining (blue) for the nuclei (A’, E’) of the same sections. B-B”, C-C”, F-F”, G-G”: High magnification images of representative nuclei of early multicellular embryos, “emb” (B-B”, F-F”), embryogenic masses, “ms” (C-C”, G-G”), microspores (D, D’) and immature zygotic embryos, “zye” (I, I’) showing 5mdC immunofluorescence in green (B, C, D,F, G, I), DAPI staining of nuclei in blue (B’, C’, F’, G’, I’) and merged images of both green and blue channels (B”, C”, F”, G”). Bars: A, A´, E, E´: 50 μm; B, B´,B´´,C, C´, C´´, F, F´, F´´, G, G’, G”, I, I’: 5 μm; D, D’: 10 μm.