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Fig. S1. Axis defects upon higher doses of HepOH. Embryos were treated with 0.02% heptanol from stage 3/4 to late neurula and processed for Pitx2c mRNA expression by WMISH at stage 25. Note the shortened axes of the two specimens on the left, and dorsal bending of the embryo on the right side.
Fig. S2. Expression of Cx40.4 mRNA at neurula and tadpole stages. Staged embryos were analyzed by WMISH using an antisense probe specific for Cx40.4. (A) Stage 18 neurula (dorsal view). Signals were present in the condensed somites, while the presomitic mesoderm (square bracket) did not express Cx40.4. (A′) Hemisection of embryo shown in (A), demonstrating absence of Cx40.4 from the GRP and adjacent tissue. (B) Stage 35 tadpole (lateral view). Persistent expression of Cx40.4 in the somites. a, anterior; d, dorsal; l, left; p, posterior; r, right; st. stage; v, ventral.
Fig. S3 Expression analysis of Cx26 and Cx32. (A) RT-PCR analysis of Cx26 and Cx32 mRNA expression from cleavage through tadpole stages. EF1α served as loading control. (B) Expression of Cx32 at stage 24. Inset shows sense control. (B′) Histological section (plane indicated by line in (B)) revealed mRNA localization in the hatching gland (hg) and stomodeal anlage (sa). a, anterior; bv, brain ventricle; cg, cement gland; d, dorsal; p, posterior; st., stage; v, ventral.
