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Fig. S2

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ZDB-IMAGE-140428-23
Source
Figures for Zhao et al., 2014
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Figure Caption

Fig. S2

Heat shock inducibility of DN-MAML-GFP, suppression of Notch signaling by DN-MAML-GFP, and cardiac regenerative deficiencies in an independent Tg(hsp70:DN-MAML-GFP) line. (A) Embryos carrying the Tg(hsp70:DN-MAML-GFP)fb10 transgene were heat shocked for 30 min at 37 °C following gastrulation (bud stage). At 24 hpf, these embryos exhibited prominent GFP fluorescence indicating DN-MAML-GFP protein expression. (B–E) Heat-shock-inducible suppression of Notch signaling in Tg(hsp70:DN-MAML) animals. Control (CTRL) and Tg(hsp70:DN-MAML) embryos were heat shocked at bud stage (B and D) or 19 hpf (E) and analyzed for gross morphological defects at 48 hpf (B and D) or relative expression of Notch target genes hey2 and her4 by qPCR at 24 hpf (E). Mind bomb (mib) mutant embryos, which are devoid of Notch signaling, were analyzed in parallel. Whereas control embryos were unaffected by heat shock (B), mib (C), and heat shocked Tg(hsp70:DN-MAML) (D) embryos displayed similar body axis curvature defects. (E) Graph showing the relative expression levels of hey2 and her4 transcripts in each experimental group. Three biological replicates were included in the qPCR analysis. Data are presented as means ± 1 SD, *P < 0.05; ***P < 0.001. (F–H) AFOG staining of representative cardiac sections at 30 dpa from heat shocked control (CTRL; F) and transgenic adults carrying an independent insertion of the Tg(hsp70:DN-MAML-GFP) transgene [Tg(hsp70:DN-MAML-GFP)fb11] (G and H). Similar to Tg(hsp70:DN-MAML)fb10 hearts, heat shocked Tg(hsp70:DN-MAML)fb11 hearts displayed prominent regenerative deficiencies as evidenced by fibrin retention (red) and collagen deposition (blue). Sections shown in G and H are from independent animals. Cardiac regeneration failed in 0 of 8 heat shocked control hearts and 5 of 5 heat shocked DN-MAMLfb11 hearts.

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