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Fig. S4
Hh signaling is upregulated in extl3/ext2 mutants and cxcl12a overexpression does not induce ectopic filopodia. Related to Figure 5-6.
(A-D) In situ hybridization of sibling and extl3/ext2 mutant embryos at 36 hpf shows the expression of the Shh target gene patched1 along the trunk (A and B) and magnified views of the notochord in the same fish in A′ and B′. patched1 stays upregulated along the tail trunk of extl3/ext2 mutants at 44 hpf (C and D), especially around the notochord (C′ and D′) where Shh signaling is active. (E-F) Tg(claudinB:GFP) fish were crossed to Tg(hsp70:sdf1a) fish and heat shocked to overexpress cxcl12a. Images were taken after heat shock (~34 hpf). A rounded primordium was observed in transgenic siblings with mild ectopic filopodia formation at the leading edge. (G-J) The Wnt/β-catenin target lef1 is expressed in the leading region of the primordium at 34 hpf (G) but a short 3 hourtreatment with the GSK inhibitor BIO (2 µM) leads to its expansion along the entire primordium (H). The chemokine receptor cxcr7b is expressed in the trailing region of the primordium at 34 hpf (I) and it is inhibited by Wnt/β-catenin signaling after BIO treatment resulting in primordium stalling (J). hpt: Hours post treatment.
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