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Figure 8. Szl-Tll Regulates FN Matrix Assembly, which Is Required to Maintain BMP Signaling(A) Embryos injected with cont-MO (20 ng), szl-MO (15 ng), cont-MO + bmp1-MO + tll1-MO (15 ng each), szl-MO + bmp1-MO + tll1-MO (15 ng each), or fn-MOs (15 ng) or treated with BMP-receptor inhibitor LDN193189 (40 μM). Confocal FN immunostaining (10 μm slice) shows FN fibrils (yellow arrow heads) between the foregut endoderm (end) and mesoderm (m), which are disrupted in szl-MO and fn-MO-injected embryos (red arrow heads). Confocal immunostaining of pSmad1 (green) and nuclei (red) in the foregut endoderm (50 μm Z-projection four to nine nuclei from the mesoderm) and in situ for bmp2, bmp4/7, and hhex indicate that BMP signaling was reduced in szl-MO, fn-MO, and LDN-treated embryos. Knockdown of Bmp1/Tll1 rescues the FN matrix, pSmad1 levels and BMP-responsive gene expression in Szl morphants. See also Figure S6.(B) Mean nuclear/cytoplasmic pSmad1 immunostaining intensity SEM in foregut cells located four to nine nuclei from the mesoderm, quantitated from 80 μm Z-projection from four sibling embryos/condition. Significantly different from contMO in t test: ∗p < 0.05, ∗∗LS mean difference > 0.3; p < 0.01; ns, not significant.(C) Mean number of activated caspase-3 foregut cells SEM. ∗p < 0.05 compared to cont-MO.Error bars reflect SEM. Image published in: Kenny AP et al. (2012) Copyright © 2012. Image reproduced with permission of the Publisher, Elsevier B. V. Experiment + Assay Source Phenotypes and Disease Xla Wt + fn1 MO + NF18-20 (immunohistochemistry) fig.8.a Anatomical Phenotype abnormal pathway-restricted SMAD protein phosphorylation Expression Phenotype decreased amount fn1.L expression in foregut Xla Wt + fn1 MO + NF18-20 (in situ hybridization) fig.8.a Expression Phenotype decreased amount bmp2.L expression in foregut decreased amount bmp4.L expression in foregut decreased amount bmp7.1.L expression in foregut decreased amount hhex.L expression in foregut Xla Wt + LDN-193189 + NF18-20 (immunohistochemistry) fig.8.a Anatomical Phenotype abnormal pathway-restricted SMAD protein phosphorylation Expression Phenotype decreased amount fn1.L expression in foregut Xla Wt + LDN-193189 + NF18-20 (in situ hybridization) fig.8.a Expression Phenotype decreased amount bmp2.L expression in alimentary system decreased amount bmp4.L expression in alimentary system decreased amount bmp7.1.L expression in alimentary system decreased amount hhex.L expression in foregut Xla Wt + szl MO + NF10.5 (in situ hybridization) fig.8.a Expression Phenotype decreased amount hhex.L expression in foregut Xla Wt + szl MO + NF18-20 (immunohistochemistry) fig.8.a Anatomical Phenotype abnormal pathway-restricted SMAD protein phosphorylation Expression Phenotype decreased amount fn1.L expression in foregut Xla Wt + szl MO + NF18-20 (in situ hybridization) fig.8.a Expression Phenotype decreased amount bmp2.L expression in foregut decreased amount bmp4.L expression in foregut decreased amount bmp7.1.L expression in foregut Larger Image Printer Friendly View

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