Fig. 7

Loss of plakoglobin increases expression of signalling genes in early stage embryos. Plakoglobin morphant embryos have increased expression of (B) bmp4, (D) bmp2b, (F) bozozok and (H) sqt compared to their respective control injected embryos (A, C, E and G). The numbers of embryos with the above phenotype were (A) 49/53 (B) 47/55, (C) 85/91 (D) 54/87 (E) 50/55 (F) 38/62(G) 61/65 (H) 49/63. (I) Semi-quantitative RT-PCR amplification from shield and sphere stage plakoglobin and control morpholino injected embryos. Lane order: 100 bp ladder, PCR cycle 13, 18, 23, 28, 33, positive plasmid control, negative template control and 1 Kb ladder. β-actin was used as a template control. Expression of bmp4, bmp2b and squint were detected five cycles and bozozok was detected ten cycles earlier in morphants than control embryos. Analysis of GFP expression in morpholino injected TOPdGFP fish. (J) In control morpholino injected fish, GFP expression is detected at the atrioventricular boundary (black arrowhead). (K) In morphant embryos, GFP is expressed throughout the heart (white arrowhead). The number of embryos with the displayed phenotype were (J) 31/39 and (K) 34/43. Co-expression study of Dkk1 and control or plakoglobin morpholino. (L) Control morpholino injected embryos at 72 hpf with normal hearts (165/165). (M) Plakoglobin morphants at 72 hpf with abnormal hearts including oedema, reflux of blood between chambers (88/149). (N) Co-injection of 20 pg of DKK1 and control morpholino resulted in oedema and reflux of blood between chambers (30/128). (O) Co-injection of 20 pg of Dkk1 and plakoglobin morpholino with normal cardiac phenotypes (10/162).