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pax6xenopus   

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Experiment details for pax6

Nicetto D et al. (2013) Assay

Suv4-20h histone methyltransferases promote neuroectodermal differentiation by silencing the pluripotency-associated Oct-25 gene.

Gene Clone Species Stages Anatomy
pax6.L laevis NF stage 29 and 30 to NF stage 33 and 34 retina , forebrain , midbrain , hindbrain

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  Figure S5. Germ layer marker gene expression in xSuv4-20h double morphants. (A) Immuno-histochemistry on ctrl-MO and xSuv4-20h double-morphant tadpoles. Panels show representative cross-sections of neural tubes stained with antibodies against the histone epitopes indicated on top. Inj injected side. Squares on double-morphant sections represent the croped pictures shown in Figure 1D. (B-F) RNA in situ hybridization analysis of ctrl-MO injected and double morphant embryos at the indicated stages using probes against Chordin, Xnr-3 and Gooscoid (B), Krox20 and Otx2 (C), Pax-6 (D), N-CAM (E), FoxD5, Geminin, Zic2, Zic3, Sox3, Sox11 and VegT (F). Chordin, Xnr-3 and Gooscoid dorsal side views; animal pole is on the top. Krox20 - dorsal views, anterior on the left. Otx2 - anterior views, dorsal on the top. Pax-6 head region; rescued embryos included. N-CAM - dorsal views of stained embryos with the anterior on the left; rescued embryos included. FoxD5, Geminin, Zic2, Zic3 - dorsal views; injected halves are lineage-traced by coinjection of LacZ mRNA and subsequent βal staining (light blue). Sox2, Sox3 - dorsal views, anterior to the left. VegT - internal stain in bisected embryos.

Gene Clone Species Stages Anatomy
pax6.L laevis NF stage 29 and 30 to NF stage 33 and 34 retina , midbrain , hindbrain , cement gland

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  Figure S8. Xenopus laevis Suv4-20h1 or h2 mRNA overexpression. (A) Overexpression of frog Suv4-20h1 and h2 enzymes causes an upregulation of H4K20me2 and H4K20me3 marks. Bulk histones from uninjected embryos or embryos bilaterally injected with increasing amounts of Suv4-20h1 or h2 mRNAs were isolated at NF11.5 and analysed by Western blot. Pan H3 antibody was used as loading control. (B, C) Morphological phenotypes of NF30-33 embryos injected with xSuv4-20h1 (B) or h2 (C) mRNA. (D) RNA In situ hybridization of NF30-33 uninjected embryos (top row) and embryos injected with Suv4-20h1 (middle row) or h2 (bottom row) mRNA using probes against Rx-1 and Pax-6. Pictures show the head of stained embryos. (E) RNA In situ hybridization analysis of uninjected embryos (top row) and embryos injected with xSuv4-20h1 (middle row) or h2 (bottom row) mRNA using probes against Ngnr-1a, Delta-like 1, N-tubulin, Xbra, MyoD, Sox17 α and Endodermin. Pictures show dorsal views of stained embryos, anterior is on the left; Xbra pictures show vegetal views of NF11 embryos; MyoD pictures show dorsal views of NF15 embryos, with the head on the left. For Sox-17 α and Endodermin sagittal sections of NF15 embryos were created; pictures show internal view of the injected halves, with anterior on the left.