DEV031161 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -

• Supplemental Table S2 -

• Supplemental Table S3 -

• Supplemental Table S4 -

• Supplemental Table S5 -

• Supplemental Figure S1 -

  Fig. S1. Time-lapse of dorsal clones. (A-H) Still images of dorsolateral clones, lateral views. Arrowheads indicate dorsal midline and arrows indicate notochord boundaries. (A-D) gfp RNA-injected control embryo from shield (A) to 90% (D). (E-H) gsc RNA-injected embryo from 60% epiboly (E) to 2-somite stage (H).

• Supplemental Figure S2 -

  Fig. S2. chd does not induce gsc. Animal views of shield (arrowhead) stage embryos stained for gsc and injected with (A) gfp (n=18) or (B) chd RNA (n=26).

• Supplemental Figure S3 -

  Fig. S3. Morpholinos. (A-F) Live chd^tt250 mutant embryos at 1 dpf; injected construct is listed in lower right. (A) control; (B) fstl1b-MO; (C) nog1-MO; (D) fstl1b- and nog1-MOs. (E,F) The most ventralized phenotypes seen in two examples of embryos injected with fstl1b-, nog1- and gsc-MOs.

• Supplemental Figure S4 -

  Fig. S4. Morpholino controls. Live embryos at 1 dpf; genotype is listed in upper right, injected construct is listed in lower right. (A-D) gsc-MO blocks dorsalization by injected gsc, indicating that endogenous gsc is required for dorsalization. (E-G) chd-MO rescued with Xenopus chd RNA. (I-L) Rescue of nog1-MO by nog1 RNA. (M-P) Rescue of fstl1b-MO by fstl1b RNA. (Q-S) Neither low nor high concentrations of standard control morpholino (equivalent to single and triple morpholino experiments, respectively) enhance the chd mutant phenotype.

• Supplemental Figure S5 -

  Fig. S5. gsc constructs in Xenopus. (A-E) Xenopus laevis embryos, anterior to the left. (A,C) Uninjected. Embryos injected dorsally with VP-gscHD (B), Xenopus gsc-VP2 (D) or gsc-VP2 RNA (E) had reduced anterior structures.