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Fig. S1 Characterization of the Tg(pax2a:GFP)e1 PPA domain. (A-A′′) Confocal projection of a Tg(pax2a:GFP)e1 embryo at 12 hpf immunostained for Pax2a. Significant overlap was seen between transgenic GFP and endogenous Pax2a; 97% of α-Pax2+ cells were GFP+ (n=380 cells from five embryos) and 87% of GFP+ cells were α-Pax2+ (n=420 cells from five embryos). (B-B22) Confocal projection of a Tg(pax2a:GFP)e1 embryo at 24 hpf co-stained with the Pax2 antibody. Few cells in the EB placodes are labeled with the transgene at this stage. Ectopic expression of transgene is seen in the acoustic and anterior lateral line ganglion (gVIII/gAll) and rhombomeres 3 and 5 (R3 and R5). (C) Confocal projection of a Tg(pax2a:GFP)e1 transgenic embryo at 12 hpf. (D) Total number of cells of the PPA, in addition to number of cell diameters along the A-P (red marker) and M-L (yellow marker) axes, were counted in 11 embryos (1186 cells total from 11 embryos) to determine variability in dimensions of this domain. Counts reveal a consistent average (Ave) with a small standard error of mean (SEM) for both total number of cells and cells across the A-P and M-L axes in the PPA of 12 hpf embryos. ov, otic vesicle; gVIII/gAll, acoustic/anterior lateral line ganglia; A-P, anterior-posterior; M-L, medial-lateral. Scale bars: 50 μm.