DEV02265 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1A
• Supplemental Figure 1B -

  Fig. S1. Nucleotide and deduced amino acid sequence of hydkk1/2/4, nvdkk1/2/4 (Fig. 1A) and nvdkk3 (Fig. 1B). Nematostella sequences represent contiguous sequences from the N. vectensis EST project and are not complete. Putative signal peptide is shaded in yellow; the red line marks the predicted cleavage site. Conserved cysteine motifs are shaded in green (CRD1) and blue (CRD2); conserved cysteine residues are in bold type.

• Supplemental Figure 2 -

  Fig. S2. hydkk1/2/4- and hywnt3a-expression in Alsterpaullone (AP)-treated animals. Polyps were incubated for 24 hours in 5 mM AP and then transferred to hydra medium. (A,B) ISH with antisense hydkk1/2/4 probe 2 days (A) and 3 days (B) after the onset of AP treatment. Note the lack of hydkk1/2/4 expression in large areas at 3 days (B). (C,D) A higher magnification image (C) and a double-ISH with hydkk1/2/4 (red) and hywnt3a (blue) (D) reveal the lack of hydkk1/2/4-expressing gland cells in these patches and hywnt3a expression in their centre (D). These expression patterns suggest that the downregulation of hydkk1/2/4 starts from the centre of the hywnt3a expression domains.

• Supplemental Figure 3 -

  Fig. S3. TOPFLASH luciferase assay in HEK293T cells. Cells were transformed with mwnt1 or hlrp6 in presence or absence of xdkk1, hydkk1/2/4 and hkremen1 (krm1). HyDkk1/2/4 potentiates canonical Wnt signaling 1.7- to 2.5-fold synergistically, when activated by Wnt1 or Lrp6. Co-injected krm1 enhances reporter activation by hydkk1/2/4/wnt1 or hydkk1/2/4/lrp6 2-fold, which is not observed in xdkk1 control (data not shown).