6533b881fe1ef96bd12d8c3d

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Additional file 10 of The Wnt-specific astacin proteinase HAS-7 restricts head organizer formation in Hydra

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Additional file 10: Fig. S8. Developmental balance between ectopic structures. (a) siHAS-7/siNdr electroporation blocks ectopic axis formation after subsequent AZK treatment. (b-c) siHAS-7/Wnt8 electroporation and AZK treatment reduces ectopic tentacle development in double axis animals (b) and leads to multiple secondary axis formation in a fraction of the treated animals (c). Red arrows denote secondary axes. The asterisk denotes the peduncle region. (d) Ectopic tentacle inhibition is clearly evident in animals electroporated with siWnt8 followed by AZK treatment. Note that few residual ectopic tentacles are detectable in c and d mostly on the side not directly hit by the electroporation pulse. (e) Continuous treatment with a low concentration of ALP (0.2 μM) leads to secondary axis development after about 3 weeks. Note that as in siHAS-7/Wnt8 treated animals, few residual ectopic tentacles are present. Representatives of at least 7 hydras examined. Scale bars = 200 μm. (f) Ratios of double and multiple axis phenotypes in hydras after electroporation with siGFP or combinations of siRNAs as indicated. AZK incubation was started 6 days after electroporation and the numbers of ectopic axes in each group were counted 12 days after AZK removal. Total numbers of animals with ectopic axis phenotype in each group were: siGFP/AZK = 0/38 (n=3), siHAS-7/siNdr/AZK = 0/63 (n=3), siHAS-7/siWnt8/AZK = 22/61 (multiple axis = 7) (n=3), Results from at least three independent experiments are shown. Each column represents the total percentage of one group, bars indicate the mean ± S.E.M. **P value