GATA4 induces the expression of ventricular and proepicardial markers in animal cap explants. (A) Ventricular gene expression is induced in animal cap explants injected with GATA4 (75%, n = 50) using WMISH analysis for the expression of myl3. (B, C) Close-up of animal cap explants shown in (A); black arrowheads are indicating myl3 expression. (D, E) Lateral and ventral views of a myl3 expression in control embryo (stage 39). (F–H) Animal caps injected with GATA4 showed incompletely overlapping expression of pan-myocardial marker tnni3 and ventricular marker myl3 (36% (n = 55), 13% showed tnni3 expression only). Similarly to the heart of control embryos (I, J), GATA4 injected AC explants show a region of overlapping expression (black arrowheads) and region expressing tnni3 alone (white arrowhead). (K) RT-PCR analysis in AC explants at stage 40 injected with GATA4 or co-injected with GATA4 and Dkk-1 mRNA showed the expression of the ventricular (myl3 and irx4) and proepicardial (tbx18) markers as in sibling control embryos at the same stage. A representative experiment out of 3 repeats is shown. a, atrium; ih, interhyoid muscle of the jaw; v, ventricle. The scale bar corresponds to 100 μm.