DEV01685 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Non-neural marker and pan-neural marker expression in Wnt8 gain-of-function experiments. Overexpression of wnt8 suppresses non-neural markers throughout the epiblast: (A) Control embryo and (B) injected embryos with wnt8 RNA 100 pg stained for p63 at 60% of epibloy. (C) Control embryo and (D) embryo injected with wnt8 RNA 100 pg stained for foxi1 at 60% of epibloy. Overexpression of wnt8 induces ectopic expression of neural markers throughout the epiblast. (E) Control embryo and (F) embryos injected with wnt8 RNA 100 pg stained for zic2.2 at 60% of epibloy. (G) Control embryo and (H) embryos injected with wnt8 RNA 100 pg stained for sox31 at 60% of epibloy. See also Fig. 6 for a similar analysis carried out using clones of wnt8 misexpressing cells.

• Supplemental Figure 2 -

  Fig. S2. wnt8 expression does not ‘self-induce’ and Wnt8 can repress otx2 in Df(Wnt8) host embryos. (A,B) Embryos containing cells derived from injected embryos with a lineage tracer (brown) and wnt8 RNA (400 pg) into the animal pole of a wild-type host embryo. (A˘-B˘˘) Close-up of transplanted cells (A˘,B˘) before biotin staining and (A˘˘,B˘˘) after biotin staining. The embryos are stained for wnt8 (blue) and otx2 (red). No wnt8 expression is observed around the transplanted clone (brown) suggesting that the repression of otx2 is caused by by Wnt8 produced by the transplanted cells (n=15/15). (C,D) Embryos containing cells derived from injected embryos with a lineage tracer (brown) and wnt8 RNA (400 pg) into the animal pole of a host embryo mutant for wnt8 (Df(Wnt8)). (C˘-D˘˘) Close up view of transplanted (C˘,D˘) before biotin staining and (C˘˘,D˘˘) after biotin staining. Wnt8 from the transplanted cells represses otx2 as no Wnt8 protein can be produced by the host embryo (mutant analysed n=10/10).