We performed conditional inhibition of ErbB signaling using AG1478, a competitive kinase inhibitor that is routinely used in zebrafish to block this signaling pathway [37 (link), 44 (link), 59 (link), 60 (link)]. A final concentration of 5 μM AG1478 (Calbiochem, San Diego, CA, USA) in 0.05 % dimethyl sulfoxide (DMSO) was added to the E3 medium containing larvae from 10 hpf until 58 hpf to inhibit SC development. Alternatively, a final concentration of 5 μM of AG1478 in 0.05 % DMSO was added to embryos between 60 hpf and 144 hpf. This treatment allows SC development but blocks ErbB signaling from 12 h before electroablation until the end of the regeneration process.
For copper treatment, CuSO4 (Merck, Darmstadt, Germany) was dissolved in E3 medium and 72 hpf larvae were exposed to 100 μM CuSO4 for 2 h and subsequently rinsed a minimum of three times in fresh E3 medium.
For copper treatment, CuSO4 (Merck, Darmstadt, Germany) was dissolved in E3 medium and 72 hpf larvae were exposed to 100 μM CuSO4 for 2 h and subsequently rinsed a minimum of three times in fresh E3 medium.
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