Fig. S6

Hyperactivation of Wnt signaling impairs zebrafish heart regeneration. Related to Figure 3. (A) Bar graph showing the relative expression levels of the Wnt target genes axin2, ccnd1 and mycn in spared tissue (blue box shown in Fig. S5A) of CTRL and Tg(DM-MAML)EC animals on 5 days post amputation (dpa) as determined by qPCR. (B) Bar graph showing the relative expression levels of axin2, ccnd1 and mycn in ventricles of wild-type animals injected intraperitoneally with DMSO, the Wnt inhibitor IWR, or the WNT activator BIO as determined by qPCR. For (A) and (B), three biological and technical replicates were analyzed. Statistical significance was determined using a Student’s t- test, n.s., not significant. **, p<0.01. *,p<05. ***,p<0.001. (C-N) Single confocal slices of sections surrounding those shown in Fig. 3B (C-F), Fig. 3C (G-J), and Fig. 3D (K-N). immunostained to detect myocardium (MF20 antibody; magenta) and counterstained with DAPI (blue). The asterisks in (K-N) highlight gaps in the myocardial wall. (O-Z) Brightfield images of sections surrounding those shown in Fig. 3E (O-R), Fig. 3F (S-V), and Fig. 3G (W-Z) stained with AFOG to detect muscle (brown), fibrin (red), and collagen (blue). The asterisks in (W-Z) highlight collagen-rich scar tissue. (AA) Box and whisker plot showing the percentages of total ventricular area occupied by scar tissue in 30 dpa wild-type animals injected with DMSO (n=8), IWR (n=7), and BIO (n=8). Scar percentage data were collected for 6-10 sections per heart and averaged to generate each data point. Statistical significance was determined using a Student’s t- test. Error bars = ± 1 SD. n.s, not significant. **, p<0.01. *,p<0.05. Scale bars=50μm.