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Gene/Clone	Species	Stage	Anatomy Item	Experimenter
sox3	xenopus

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Experiment details for sox3

Klisch TJ et al. (2006) Assay

Mxi1 is essential for neurogenesis in Xenopus and acts by bridging the pan-neural and proneural genes.

Gene	Clone	Species	Stages	Anatomy
sox3.S		laevis	NF stage 13	trigeminal placode , neural plate

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Fig. 8. Xmxi1 induces proliferation in the open neural plate of Xenopus embryos. MT-Xmxi1 (500 pg), EnR-Xmxi1 (500 pg), and Xmxi1-GR (500 pg, induced at stage 10.5 with dexamethasone), Xmxi1-MO (12.5 ng), or the control-MO (Co-Mo) (12.5 ng) was injected in Xenopus embryos. Proliferation on the injected side compared with the noninjected side was measured by counting (A) pH3 or (B) BrdU-positive cells of 15 consecutive sections of five embryos in the open neural plate region. Shown is the average number of positive cells per section. Error bars indicate the standard error of the mean. (C) Forced cell cycle arrest does not alter the phenotype of Xmxi1-injected embryos. In Xmxi1-GR-injected embryos, proliferation was blocked by HUA treatment at stage 10 and induced at stage 10.5 with dexamethasone. (D) Inhibition of MT-XMxi1-induced apoptosis does not rescue loss of N-tubulin. Overexpression of MT-Xmxi1 (500 pg) led to an increase in apoptosis as seen by TUNEL staining (upper panel, right). Coinjection of human Bcl2 mRNA (500 pg) blocked apoptosis (lower panel, right) but did not influence the effects of MT-Xmxi1 on Sox3 and N-tubulin.

Gene	Clone	Species	Stages	Anatomy
sox3		xenopus	NF stage 14	trigeminal placode , neural plate

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Fig. 5. Xmxi1 activates ectopic Sox3 and inhibits neuronal differentiation. (A–N) Whole-mount in situ hybridization of stage 14 embryos injected with 500 pg of MT-Xmxi1, MT-Xmad1, MT-Xmxi1-DBM, EnR-Xmxi1, or Max, as indicated in the upper right hand corner. Antisense probes used are indicated in the lower left hand corner. The injected side is always on the right, and all embryos are shown as dorsal views, anterior down with the exception of C and N, which are ventral views.

Gene	Clone	Species	Stages	Anatomy
sox3.S		laevis	NF stage 14	pre-chordal neural plate , chordal neural plate , neural plate , anterior neural fold

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Fig. 1. (A) Comparison of the predicted amino acid sequence of Xmxi1 with the mouse Mxi1 isoforms. GenBank accession number Xmxi1: DQ137875. (B) RT-PCR analysis of Xmxi1 at different stages of development. Histone H4 was used as a loading control. (C) Whole-mount in situ hybridization expression analysis of Xmxi1 in comparison with X-ngnr-1. (D–F) Double labeling of Xmxi1 and Sox3 expression. Sox3, red; Xmxi1, dark blue. (D) Stage 14, anterior view. (E) Stage 14, dorsal view. White arrowhead marks the lateral stripe of Xmxi1. (F) Transversal section of E. (G–N) Spatial expression of Xmxi1 during later stages of Xenopus development. (G) Stage 13, dorsal, anterior up. (H) Stage 16, dorsal view, anterior left. (I) S1 transversal section. (J) Stage 22, lateral. (K) Stage 27, anterior (L) Stage 37, lateral. (M) S2 transversal section. (N) S3 transversal section, dark brown staining at the roof of the spinal chord is pigmentation not Xmxi1 staining. el, epithelial layer; hb, hindbrain; i, intermediate; iz, intermediate zone; l, lateral; le, lens; m, medial; mb, midbrain; n, notochord; op, olfactory placode, r, retina; sl, sensory layer; st, stage; tg, trigeminal placodes; vz, ventricular zone.

Gene	Clone	Species	Stages	Anatomy
sox3		xenopus	NF stage 24	brain , midbrain , hindbrain , spinal cord

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Fig. 7. (A) Xmxi1 induces neuronal differentiation in neuralized late, but not early animal caps. Animal caps were harvested from embryos injected with noggin (50 pg) and MT-Xmxi1 (500 pg), as indicated. At the equivalent of stage 14 or 20, the caps were harvested and analyzed by real-time RT-PCR. Expression levels were normalized to ODC and compared to control caps, which were set to 1. (B–E) The inhibition of neuronal differentiation by Xmxi1 is only transient. At tailbud stages, MT-Xmxi1 (750 pg) ectopic Sox3 is still present (B) and ectopic N-tubulin expression is also detected (D). (C, E) Transversal sections of B and D, respectively. Red arrowhead marks ectopic expression.