Fig. 8

New vPNS genes are regulated by BMP and Notch signaling pathways in B. lanceolatum embryos. A–L In situ hybridization for candidate genes expressed in ventral epidermis: Wnt11 at early neurula (A–C), HairyE (D–F), Gata4/5/6 (G–I) and Lbx1/2 (J–L) at mid neurula stages in control embryos (A, D, G, J), following treatment from late gastrula stage (12 hpf) with BMP4 protein (B, E, H, K) or with dorsomorphin (C, F, I, L). M–B’ In situ hybridization for candidate genes expressed in ESNs: Nhlh (M–P), BLAG19000137 (Q–T), Islet (U–X) and Brsk (Y–B’) at late neurula stages in control embryos (M, Q, U, Y), following treatment from late gastrula stage (12 hpf) with BMP4 protein (N, R, V, Z), with DAPT (O, S, W, A’) or a combination of BMP4 and DAPT (P, T, X, B’). Embryos are shown in lateral view with dorsal to the top and anterior to the left. Black arrowheads indicate ESNs when not clearly visible and arrows indicate the anterior most epidermis of the embryo presenting an accumulation of ectopic ESNs. Each experiment has been performed twice. The number of embryos examined for each condition is between 15 and 40, and the pictures shown correspond to the phenotype observed in over 90% of the embryos. Scale bar: 50 μm