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Supplementary Material

DEV046631 Supplementary Material

Files in this Data Supplement:

  • Supplemental Figure S1 -

    Fig. S1. Hydractinia echinata. (A) Life cycle. The arrow in the fertilization image indicates sperm. (B) Colony structure.

  • Supplemental Figure S2 -

    Fig. S2. Alignment of Tcf and β-catenin cDNAs from various animals. (A) Alignment of Tcf genes from Hydra magnipapillata (HyMtcf, accn XP_002159974), H. vulgaris (HyVtcf, accn AAG13664.1), Hydractinia echinata (HeTcf), Nematostella vecternsis (NvTcf, accn ABF55257.1), and human Tcf1, 4 and 7 (accns CAA87440.1, CAB97214.1 and EAW62279.1, respectively). (B) Alignment of β-catenin from Hydra vulgaris (accn AAQ02885.1), H. magnipapillata (accn XP_002159953.1), Podocoryne carnea (accn ABI74628.1), Hydractinia echinata and Homo sapiens (accn BAG70078).

  • Supplemental Figure S3 -

    Fig. S3. Additional phenotype images of various experiments. (A-C) Metamorphosis treatments. (A) Rescued animal 24 hours post-metamorphosis showing normal polyp phenotype after joint azakenpaullone and Tcf RNAi treatment. (B) An azakenpaullone-treated animal 24 hours post-metamorphosis. Ectopic tentacle buds appear at the same time as normal tentacle buds in control animals. Aboral structures fail to develop in azakenpaullone-treated animals. (C) An azakenpaullone-treated metamorphosed animal, 2 days after induction of metamorphosis. Tentacle buds develop into fully functional tentacles, aboral structures are still absent. (D-F) Regeneration treatments. (D) Cut polyp body column, 9 days after the start of a 48-hour treatment with azakenpaullone. All tissue has been converted to an oral fate. The animal now merely consists of three joined regenerated tentacles. (E,F) Similarity between LiCl and late-stage Wnt3 RNAi-treated polyps. (E) Polyp cut from the colony at the base then treated with 28 mM LiCl. Three heads (top) developed side by side at the cut (originally aboral) end with the original head still visible at the bottom of the image. (F) Wnt3 RNAi polyp showing side-by-side head regeneration similar to that of the LiCl treatment. RNAi polyp cut from the colony at the base then head removed. Head regeneration was initially inhibited, then, as the effect of the RNAi began to fade, two heads regenerated side-by-side. (G-I) Stolon development in a Wnt3 RNAi-treated polyp. The three images are of the same polyp. (G) Three days after cutting, a stolon bud (arrow) formed but there was no head regeneration. (H) Five days after cutting, the stolon has attached to the substratum (arrow), facilitating its growth. A head has regenerated at the same end of the polyp as the stolon. (I) Nine days after cutting, the stolon has branched multiple times and continues to grow. Scale bars: 200 µm.