Fig. S3

(related to Figure 4). Ajuba proteins regulate the number of Isl1-expressing cells in the heart

(A) Number of Isl1⁺ cells at the venous pole in and at the periphery of the heart at different developmental stages. (B) Number of Isl1⁺ cells at the venous pole in and at the periphery of the heart in control and Ajuba morphant embryos at 30 and 48 hpf. n = 3; *p<0.05; **p<0.01. (C) Number of Isl1⁺ cardiomyocytes at the venous pole in control and Ajuba morphant embryos at 48 hpf. n = 3; **p<0.01. For these experiments (A, B, C), confocal images of control and Ajuba-morphant Tg(myl7:EGFP-HsHRAS)^s883 embryos stained with anti-Isl1 antibodies (red) and visualized by GFP fluorescence (green), were projected using ImageJ and Isl1⁺ cells or Isl1⁺ cardiomyocytes were counted. (D) In situ hybridization for Isl1 of control, LIMD1 and WTIP morphant embryos at 48 hpf. (E) Total number of cardiomyocytes in control, Ajuba, Isl1 and Isl1/Ajuba-deficient zebrafish embryos at 48 hpf. n = 3; ***p<0.001. (F) In situ hybridization for Isl1 of control, mIsl1 and mIsl1ΔLIM2 mRNA injected embryos at 48 hpf.