XB-IMG-81296
Xenbase Image ID: 81296
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Figure 5. Szl Is Required to Maintain Robust BMP Signaling in the Foregut(A) Western blot of pSmad1/5/8 and total Smad1 in foregut explants from cont-MO and szl-MO embryos at different stages shows reduced BMP signaling in Szl morphants.(B) Szl-depleted embryos exhibit reduced pSmad1 signaling in the foregut. Fifty micrometer confocal Z-projections of pSmad1/5/8 (green) and nuclei (red) in stage 20 sibling embryos injected with cont-MO (20 ng), szl-MO (15 ng), or bmp2-MO (20 ng) or treated with BMP-receptor inhibitor LDN193189 (40 μM). Yellow dashed lines indicate the mesoderm-endoderm boundary. Dashed white lines in nuclear channel of the cont-MO illustrate the subregion of the foregut, based on number of nuclei from the mesoderm. See also Figure S3.(C) Szl- and Bmp2-depleted embryos have reduced pSmad1 levels in the deep foregut endoderm. Mean nuclear/cytoplasmic ratio of pSmad1 immunostaining intensity SEM in foregut cells at different distances from the mesoderm, quantitated from 80 μm Z-projections from four sibling embryos/condition. ∗Significantly different from szl-MO, p < 0.05.(D) Expression of bmp2, bmp4/7, and BMP-target genes szl, vent1/2, and hhex were downregulated by injection of the szl-MO (15 ng) and rescued by subsequent injection of recombinant BMP2 protein (4 ng) into the foregut at stage 12.(E) Mean number of activated caspase-3 positive foregut cells SEM (n = 5 embryos / condition), ∗p < 0.01 in Student t tests.Error bars reflect SEM. Image published in: Kenny AP et al. (2012) Copyright © 2012. Image reproduced with permission of the Publisher, Elsevier B. V.
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