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Fig. S1

Further phenotypic analysis of lyc1. Related to Figure 1

(A-B) Representative overall morphology of WT and lyc1 mutant embryonic hearts and pericardial cavities at 3 (A) and 5 dpf (B). Double-sided arrows indicate the distance between the pericardium and heart. Dashed line indicates the outline of the myocardium. (C) In situ hybridization analysis of flt4 (n=22), dab2 (n=16), coup-TFII (n=20), ephrinb2a (n=17) and vegf-c (n=28) in lyc1 mutant embryos revealed no alterations in normal expression patterns. N values indicate the total number of embryos examined from an incross of known heterozygotes (expected 25% lyc1 mutants). Individual genotype confirmed mutant embryos are shown in the right hand panels. (D) Expression of cxcr4a was unchanged at 32 hpf in control WT/MO-pkd1b (embryos show no phenotype after pkd1b knockdown only and internally control for MO toxicity) (n=39/40) compared with phenotypically mutant (based on body curvature in the presence of MO-pkd1b) lyc1/MO-pkd1b embryos (5ng MO) (n=16/16). Expression of cxcr4b was unchanged at 32 hpf in control (WT/MO-pkd1b) (n=36/38) compared with phenotypically mutant lyc1/MO-pkd1b embryos (5ng MO) (n=7/7). Expression of cxcl12a was unchanged at 32 hpf in control (WT/MO-pkd1b) (n=50/52) compared with phenotypically mutant lyc1/MO-pkd1b embryos (5ng MO) (n=14/14). Expression of cxcl12b was unchanged at 32 hpf in control (WT/MO-pkd1b) (n=28/37) compared with phenotypically mutant lyc1/MO-pkd1b embryos (5ng MO) (n=6/7). (E) Expression of klf2a at 24 (n=10/10) and 32 hpf (n=9/10) is normal in WT embryos. Expression of klf2a at 24 hpf (n=14/14) and 32 hpf (n=19/20) in lyc1 mutant embryos.

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