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Fig. S3
Pharmacologically enhancing RA levels can recapitulate Cyp26-deficient heart defects, while inhibiting RA production and co-injecting cyp26a1 mRNA can restore Cyp26-deficient heart defects.
(A,B) IHC of DMSO or Ketoconazole (Keto) treated embryos at 48 hpf with ventricle in red (MHC) and atrium in green (AMHC). (C) Graph depicting ventricular counts of DMSO or Ketoconazole treated embryos at 48 hpf. (D,E) Hearts of Tg(myl7:EGFP) embryos at 72 hpf that were treated with DMSO or 2 μM RA beginning at 24 hpf. Arrows denote ectopic cardiomyocytes. (F) Graph of ventricular counts embryos at 48 and 72 hpf (n = 10 per group) of DMSO or 2 μM RA treatment beginning at 24 hpf. (G-J) Hearts at 48 hpf in control or Cyp26-deficient Tg(myl7:EGFP) embryos treated with DMSO or DEAB. (K) Graph depicting ventricular cell counts at 48 hpf for DMSO, DEAB, Cyp26-deficient + DMSO, and Cyp26-deficient + DEAB treated embryos. (L) Graph depicting ventricular cell counts at 48 hpf in control and Cyp26-deficient Tg(myl7:DsRed-NLS) embryos injected with cyp26a1 mRNA. (M-P) Hearts at 48 hpf from control, cyp26a1 mRNA-injected, Cyp26-deficient and Cyp26-deficient injected with cyp26a1 mRNA Tg(myl7:EGFP) embryos. Error bars are SEM, asterisks denote p<0.05 by Student’s t-test. All images are frontal views with anterior up; n>20 embryos for (A,B,D,E,G-J,M-P). Scale bars: 50 mm.