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smad1xenopus nucleus 

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Experiment details for smad1

Cibois M et al. (2015) Assay

BMP signalling controls the construction of vertebrate mucociliary epithelia.

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Gene Clone Species Stages Anatomy
smad1.L laevis NF stage 9 nucleus
smad1.L laevis NF stage 10 nucleus
smad1.L laevis NF stage 12 nucleus
smad1.L laevis NF stage 15 nucleus
smad1.L laevis NF stage 19 nucleus

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  Fig. 1. BMP pathway activity in the developing Xenopus epidermis. (A) Immunostaining of cryosectioned Xenopus embryos revealed that the pSmad1/5/8 signal (red) colocalises with nuclear DAPI staining (blue) in both inner and outer layer cells. (B) (Left) In embryos injected with linearised plasmid DNA for the MCC reporter α-tub::GFP (50 pg/embryo), the pSmad1/5/8 signal (red) colocalised with GFP immunostaining (green). (Middle) In embryos injected with linearised plasmid DNA for the ionocyte reporter pendrin::GFP (25 pg/ embryo), the pSmad1/5/8 signal (red) colocalised with GFP immunostaining (green). (Right) The pSmad1/5/8 signal (red) colocalised with P63 protein expression (white). For each marker and stage, a representative cell (yellow arrowhead) is displayed at higher magnification to show the double staining. The white dashed and dotted lines respectively mark the apical surface of the ectodermal outer layer and the boundary between the outer and the inner ectodermal layers.

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Gene Clone Species Stages Anatomy
smad1.L laevis NF stage 9 to NF stage 10 nucleus

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  Figure S1: Validation of pSmad1/5/8 staining in response to BMP manipulation. (A): Injection of stage 9 Xenopus embryos with recombinant BMP4 led to increased Smad1/5/8 phosphorylation and nuclear localization. (B): The percentage of nuclei positive for the pSmad signal is significantly higher in BMP-treated than in control embryos (Student's t-test). (C): Knocking down BMP2, BMP4 and BMP7 with specific morpholino oligonucleotides inhibited Smad1/5/8 phosphorylation. Embryos were injected with either GFP mRNA alone (upper row) or GFP mRNA and the BMP2/4/7 MOs (lower row). The pSmad1/5/8 signal was lost in cells that received the BMP2/4/7 MOs, but not in those that received only the GFP mRNA.