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Experiment details for msx1

Rankin SA et al. (2012) Assay

Suppression of Bmp4 signaling by the zinc-finger repressors Osr1 and Osr2 is required for Wnt/β-catenin-mediated lung specification in Xenopus.

Gene Clone Species Stages Anatomy
msx1.L laevis NF stage 33 and 34 to NF stage 35 and 36 otic vesicle , brain , spinal cord , eye , mouth primordium , [+]

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  Fig. 7. Osr1/Osr2 can repress bmp4 expression. (A) GR-mOsr1 RNA (750 pg) was injected into both dorsal-vegetal cells at the eight-cell stage and embryos were dexamethasone treated at stage 20 to induce the construct and analyzed at stage 34/35 for expression of the indicated genes. Insets show ventral views of wnt2b expression. (B) The X. laevis bmp4 promoter construct, showing two putative odd-skipped-binding sites. Green sequence indicates consensus odd skipped-binding site, red sequence is the exact bmp4 promoter sequence, and mutations to these sites are shown below indicated by ‘δ’ and in black. (C) RNA (750 pg) encoding Xenopus, mouse or Drosophila Osr factors was co-injected with the –2123 bmp4: luciferase reporter at the 32-cell stage and luciferase activity was assayed at stage 12. Average relative activity ±s.d. (D) Xenopus Osr1 or Osr2 RNA (750 pg) was co-injected with the wild-type bmp4 reporter or the mutated reporter constructs indicated in B, and the resulting average fold repression ±s.d., of the luciferase reporter from three independent experiments was determined. *P<0.005.

Gene Clone Species Stages Anatomy
msx1.L laevis NF stage 35 and 36 lens , brain , spinal cord , epidermis , cement gland , [+]

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  Fig. 6. Osr1/Osr2 promotes lung specification and growth by restricting BMP signaling. (A). Analysis of Osr1/Osr2-MO and control MM-MO stage 35 embryos indicated that expression of bmp4 and BMP-target genes msx1 and hand1 are expanded into the ‘wnt2b-domain’ of the lpm on the Osr1/Osr2-depleted side. Black arrows indicate normal expression, red arrows indicate absent expression and green arrows indicate enhanced expression. (B) Confocal immunostaining of stage 35 embryos for phospho-Smad1/5/8 (green) and fibronectin (red). Scale bars: 50 μm. White arrows in B indicate ectopic pSMAD1 expression in dorsal foregut endoderm. (C) Analysis of nxk2.1, sox2, wnt2b and raldh2 at stage 35 or sftpc at stage 41 in embryos injected with Osr1/Osr2-MOs or MM-MOs and treated at stages 25-35 with either DMSO (vehicle control) or 10 μM LDN193189. Insets show ventral views of wnt2b expression. For analysis of sftpc expression at stage 41, embryos were removed from the LDN193189 at stage 35 and further cultured in DMSO vehicle until stage 41. Black arrows indicate normal expression, red arrows indicate absent expression, green arrows indicate expanded expression and white arrows indicate rescued expression.