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nkx2-1xenopus   

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Experiment details for nkx2-1

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
nkx2-1.L laevis NF stage 28 to NF stage 32 brain , forebrain , midbrain , hindbrain , endoderm , [+]
nkx2-1.L laevis NF stage 35 and 36 brain , endoderm , eye , thyroid primordium , lung primordium

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  Fig. 4. FGF, Wnt/β-catenin and RA signaling regulate Xenopus lung development in a pathway with Osr1/Osr2. (A-D) Embryos were treated from stages 25-35 with DMSO vehicle (A), 100 μM PD173074 (B), 50 μM XAV939 (C) or 10 μM BMS493 (D), and assayed at stage 35 with the indicated probes. (E). Embryos were treated between stages 28 and 35 with vehicle (DMSO), 10 μM BIO, 80 μM SU5402, a combination of 80 μM SU5402 + 10 μM BIO, 10 μg/ml cycloheximide (CHX) or CHX +10 μM BIO, and analyzed for nkx2.1 at stage 35 and sftpc at stage 40. Black arrows indicate normal expression, red arrows indicate absent expression, yellow arrows indicate reduced expression and green arrows indicate expanded or rescued expression.

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 32 to NF stage 35 and 36 brain , forebrain , endoderm , pharyngeal arch , eye , [+]

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  Fig. 5. Osr1/Osr2 act upstream of Wnt-mediated pulmonary specification and RA-regulated lung bud growth. (A-G) Osr1/Osr2-MO- and MM-MO-injected embryos were treated at stages 25-35 with DMSO vehicle, 10 μM BIO or 1 μM all-trans-retinoic acid (RA), and analyzed for nkx2.1, sox2, wnt2b and raldh2 at stage 35 (A-F) and sftpc at stage 42 (dorsal view) (G). Black arrows indicate normal expression, red arrows show absent expression, green arrows and lines indicate expanded expression and white arrows indicate rescued expression.

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 33 and 34 to NF stage 35 and 36 brain , forebrain , endoderm , trachea , lung , [+]

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  Fig. 3. Osr1/Osr2 are required for wnt2b and raldh2 expression in the lpm. (A-X) Analysis of stage 34/35 unilateral-injected embryos shows that Osr1/Osr2 are required for expression of nkx2.1 (A-C) and sox2 (D-F) in the epithelium, whereas shh (G-I) is unaffected. In the lpm, wnt2b (J-L) and raldh2 (M-O) are absent, while foxf1 is unaffected (P-R). Black arrows indicate normal expression on the control side and red arrows indicate lack of expression on the Osr1/Osr2-MO-injected side (right). Endoderm outlined by a yellow line. Numbers of embryos with the shown phenotype are indicated. fg, foregut; hg, hindgut; de, dorsal endoderm; pe, posterior endoderm; e, esophagus; l/t, presumptive lung and trachea; lv, liver; lpm, lateral plate mesoderm; k, kidney; s, stomach.

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 33 and 34 to NF stage 35 and 36

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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
nkx2-1.L laevis NF stage 37 and 38 brain , trachea , lung , eye , foregut , [+]

  nkx2.1 (NK2 homeobox 1) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage st 37 & 38. top panel: lateral view, anterior left, dorsal up, and sections M' and M" through foregut region as indicated.

Gene Clone Species Stages Anatomy
nkx2-1.L laevis NF stage 42 endoderm , trachea , lung , lung primordium

  Confocal immunoflurescence of Nkx2-1 AB1 (green) staining trachea and lung buds/lung primordia, along with Sox2 AB1 (red) staining stomach and esophagus, in a NF stage 42 X. laevis embryo.