Morpholino-mediated knockdown of pbx1 disrupts swim bladder, pancreas, and pharyngeal arches development. A,B: Swim bladder development in 6-dpf gut GFP transgenic embryo injected with StdMO (A) and Pbx1MO (B) showing non-inflated swim bladder in pbx1 morphants. C-H: In situ hybridization of swim bladder markers prdc, foxA3, and anxa5 using embryos injected with StdMO and Pbx1MO at 36 (C, D), 48 (E,F), and 60 hpf (G,H). I,J: In situ hybridization for anxa5 in 60-hpf embryos injected with Pbx1-Ex2MM (I) and Pbx1-Ex2MO (J). K,L: Expression of pbx1 in mutants deficient in hedgehog signaling. Reduced pbx1 expression in the syu mutant at 72 hpf (K) and absence of expression of pbx1 in the swim bladder of smu mutant at 72 hpf (L). M-P: Splice-blocking morpholino-mediated knockdown of pbx2 or pbx4 disrupts swim bladder development. In situ hybridization for anxa5 in 60-hpf embryos injected with Pbx2-Ex4MM (M) and Pbx2-Ex4MO (N), Pbx4-Ex3MM (O), and Pbx4-Ex3MO (P). Q,R: In situ hybridization for pancreas marker, pdx-1, in 36-hpf embryos injected with StdMO (Q) and Pbx1MO (R). S,T: In situ hybridization for dlx2a, a pharyngeal arches marker in 36-hpf embryos injected with StdMO (S) and Pbx1MO (T). Red arrow, swim bladder; black arrow, pancreas; blue arrow, posterior pharyngeal arches.
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