DEV002352 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Temporal dependency of putative direct targets of MEK/ERK, Notch and Nodal signalling pathways to pharmacological inhibitors of the three pathways. Graphs show percentage (y-axis) of embryos with strong (orange) or weak (yellow) expression. On the x-axis, C indicates control embryos; numbers indicate the time (in minutes) that the embryo was placed in inhibitor prior to fixation. n indicates the minimum number of embryos analysed for each time point. For each treatment, two independent experiments are shown. Scoring was carried out as follows: Ci-Otx, 'strong' means 3-4 cells positive, 'weak' means 1-2 cells positive; Ci-Hesb, 'strong' means at least one A8.16 or B8.6 blastomere with strong expression, 'weak' means at least one cell with weak expression; Ci-Delta2, trunk lateral cells gave the most robust expression at this stage, so only expression in these cells was scored, 'strong' means at least one trunk lateral cell with strong expression, 'weak' means at least one trunk lateral cell with reduced expression.

• Supplemental Figure S2 -

  Fig. S2. Expression of various neural plate markers following Delta2/Notch inhibition. Marker analysed is indicated on the left and embryo treatment is indicated above the panels. For medial markers, numbers indicate the mean number of cells (c) per embryo expressing a given gene, n=total number of embryos analysed. Similar results were observed with dnDel2 injection (Ci-Mnx, 3.3c, n=21). For Ci-Tbx6b, the graph shows the percentage of embryos with strong or weak expression following the key in Fig. 5, with scoring the same as for Ci-MRF (Fig. 5).