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Fig. S3 Conditional, spatially controlled induction using additional TetA-GBD drivers and TetRE responders. (A) Transgenic driver construct for expression of TetA-GBD Cherry specifically in the myocardium. (B) Transgenic driver construct for expression of TetA-GBD Cherry specifically in her4.1 expression domains, primarily in the CNS. (C) her4.1:TetA-GBD Cherry drives efficient induction of Axin1-YFP in embryos doubly transgenic with TetRE:Axin1-YFP. Embryos were treated with EtOH or 25 μg/mL Dox plus 100 μM Dex from 24 h postfertilization (hpf) and photographed at 48 hpf. (D) Transgenic responder line construct for expression of Dkk1-GFP. (E) Heart-specific expression of Dkk1-GFP in myl7:TetA-GBD Cherry; TetRE:Dkk1-GFP double transgenic fish. Images of double transgenic embryos at 72 hpf, treated with EtOH vehicle or 25 μg/mL Dox plus 100 μM Dex from 48 hpf. Note that secreted Dkk1-GFP protein accumulates in the pericardial sac (arrow) in Dox/Dex-treated embryos. *Yolk background fluorescence. n = 20 EtOH, 24 Dox/Dex. (F) Wnt/β-catenin loss-of-function phenotypes as evidenced by posterior truncations and expanded eyes (arrow) in TetRE:Dkk1-GFP embryos injected with 95 pg TetA-GBD RNA and treated with 10 μg/mL Dox plus 100 μM Dex from 4 hpf until 24 hpf. n = 7 EtOH, 8 Dox/Dex.