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Gene/Clone	Species	Stage	Anatomy Item	Experimenter
foxe3	xenopus

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Experiment details for foxe3

Murato Y and Hashimoto C (2009) Assay

Xhairy2 functions in Xenopus lens development by regulating p27(xic1) expression.

Gene	Clone	Species	Stages	Anatomy
foxe3.L		laevis	NF stage 15	ectoderm , cement gland primordium , ventral , anterior

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Figure 4. Xhairy2 knockdown reduces the expression of LF marker genes in neurulae without marked changes in PPE marker expression. Micro-injection and lineage tracing were performed as described in Figure 3. Injected MO is indicated at the upper right corner and WISH probe is indicated at the lower right corner of each panel. a: Schematic presentation of LF in neurulae. b-i: LF marker genes. b, c: Two-color WISH of st.-15 embryos to confirm LF position. LF expression (black arrowheads) of pax6 (magenta) and six3 (purple) is indeed outside of the neural plate border that is marked by Xhairy2 (light blue). j, k: Embryos were injected with 6.9 ng of Xhairy2 MO and/or 20 pg of Xhairy2b mRNA where Xhairy2 MO does not bind in order to check the specificity of Xhairy2 MO to LF-loss phenotypes. I: Quantitative summary of j and k. Error bar: standard deviation of 5 independent experiments. m-t: PPE marker genes. Red arrowheads indicate reduction of marker gene expression. All the samples were pictured from the anterior side with the dorsal side up.

Gene	Clone	Species	Stages	Anatomy

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Figure 6. Xhairy2 knockdown affects proliferation within LF at late neurula stage, but not at mid-neurula stage. Micro-injection was performed as described in Figure 3. MO- or mRNA-injected embryos were further injected with BrdU and incubated for 1 hr prior to fixation at st. 16 (a-d) or st. 19 (e-h). To visualize LF, samples were stained with pax6 (a-d) or foxE3 (e-h) before BrdU detection. a-h: Whole-mount view from the anterior side with the dorsal side up. Red arrowheads indicate reduction of marker gene expression. a'-h': Horizontal sections of the corresponding samples for counting BrdU-positive cells within LF that is defined by LF expression of pax6 (a'-d') or foxE3 (e'-h'). Anterior side up. Red rectangles indicate LF or the corresponding region without marker gene expression. i: Quantitative summary of BrdU count. Five successive sections from each sample were subjected to counting of BrdU-positive cells within LF. The number of BrdU-positive cells from each section was summed per sample. Then, the proportion of BrdU-positive cells of the injected side with respect to the un-injected side was calculated per sample. The mean value of 3 independent samples is presented as the representative value. Error bar: standard deviation (n = 3).

Gene	Clone	Species	Stages	Anatomy
foxe3		xenopus	NF stage 28	lens , eye , lens placode

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Figure 3. Effects of Xhairy2 knockdown on the expression of lens and retinal marker genes in tail-bud-stage embryos. Co MO (3.4 ng: a-f') or Xhairy2 MO (6.9 ng: g-l'), together with EYFP and lacZ mRNA, was micro-injected into one dorsal animal blastomere at the 8-cell stage, and the embryos were harvested at st. 25 (for six6), st. 28 (for notch2, foxE3, L-maf, gamma 1-cry), or st. 32 (for rx1), followed by WISH. MO-receiving cells were visualized by X-gal staining (green). Red arrowheads indicate reduction of marker gene expression. Note that Xhairy2 knockdown reduced the expression of LP marker genes, while virtually no effect was seen on the expression of retinal marker genes six6 and rx1.