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Fig. 6 Regulation of the Otp protein by PACAP/PAC1 signaling pathway. (A-H) Embryos were injected with either a control pac1 missense morpholino (Control, A,B), or with antisense morpholinos directed against pac1 (pac1MO, C,D), otpa (otpaMO, E,F) or combination of pac1+otpa (pac1MO+otpaMO, G,H) and were thereafter (52 hpf) subjected to either in situ hybridization with an otpb-directed probe (A,C,E,G), or to immunohistochemistry with an anti-Otp antibody (B,D,F,H). Inset in panel D shows western blot analysis of nuclear-enriched protein extracts from control and pac1MO-injected embryos using antibodies directed to either Otp or the nuclear protein Rcc1 (internal protein loading control). The activity of each morpholino was evaluated by anti-tyrosine hydroxylase (TH) immunostaining. Scale bars: 100 μm in A,C,E,F; 50 μm in B,D,F,H.