Xenbase Image ID: 150107

Figure S2: Interfering with the BMP pathway alters the numbers of MCCs and ionocytes in the developing Xenopus epidermis. (A-D): 8-cell stage embryos were injected into the animal ventral blastomeres with synthetic mRNAs coding for GFP, alone (control; A, B, C) or together with mRNAs coding for a constitutively active (CA-Alk3; A', B', C') or a dominant negative (DN-Alk3; A'', B'', C'') version of the BMP receptor ALK3, then hybridized at stage 25 with a probe against the ionocyte marker v1a and antibodies against acetylated tubulin and against GFP. Activation of the BMP pathway by CA-Alk3 injection resulted in a decrease in the numbers of MCCs and Development | Supplementary Material Development 142: doi:10.1242/dev.118679: Supplementary Material ionocytes. Inhibition of the BMP pathway by DN-Alk3 injection results in an increase in the numbers of MCCs and ionocytes. (D): quantification of the different cellular populations in injected epidermal clones. The bars represent the total number of GFP positive cells scored. Magenta: acetylated tubulin positive MCCs; red: v1a positive ionocytes; green: GFP positive cells negative for both acetylated tubulin and v1a. (E): Embryos injected with a synthetic mRNA coding for a dominant negative form of the zebrafish BMP pathway nuclear effector Smad5 (dnSmad5), showed supernumerary -tubulin positive MCC precursors, which only partially managed to intercalate. (F-H): Embryos were injected in the blastocoele with BSA (control) or recombinant BMP4 protein (BMP4), fixed at stage 10, cryosectioned and immunostained with an antibody against phosphorylated histone H3 (red), a hallmark of cells in mitosis. DAPI (blue) was used to stain the nuclei. The graph in (H) shows the ratio of phospho-H3 positive nuclei to the total number of ectodermal DAPI stained nuclei. BMP4 injection did not significantly modify the number of mitotic nuclei compared to controls (Student's t-test). In F-G', the animal pole (ectoderm) is up. Image published in: Cibois M et al. (2015) Copyright © 2015. Image reproduced with permission of the Publisher. Gene Synonyms Species Stage(s) Tissue h3-3a.L h3.3, h3.3a, h3f3a X. laevis Throughout NF stage 10 embryo nucleus tuba4a.L alpha tubulin, alpha-tubulin, tuba4, tuba4b X. laevis Throughout NF stage 25 epidermis multiciliated cell ciliated epidermal cell atp6v1a.L atp6a1, atp6v1a1, atpv1a, ho68, v1a, va68, vma1, vpp2 X. laevis Throughout NF stage 25 epidermis ionocyte Image source: Published Larger Image Printer Friendly View

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