Immunofluorescence Protocol for Zebrafish Larvae

Immunofluorescence was performed as previously described (Cronan et al., 2016 (link)). Briefly, zebrafish larvae were fixed in Dent’s fixative overnight at 4°C, rehydrated in PBS containing 0.5% tween 20, and then blocked for 1 h in PBDTxGs (PBS containing 1% BSA, 1% DMSO, 0.1% Triton X-100, 2% goat serum). Mouse anti-E-cadherin antibody, clone 36 (BD, 610181) was added at a 1/500 dilution followed by incubation overnight at 4°C. Larvae were washed in PBDTxGs and then Alexa Fluor 647 Goat Anti-Mouse IgG (H+L) antibody (ThermoFisher, A-21236) added at a 1/500 dilution followed by incubation overnight. Larvae were washed 5 times in PBDTxGs before analysis.