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hand2 and dlx3b/4b/5a repress dorsal identity. (A-T) Projections of confocal stacks of 36 hpf RNA in situs. Arch one and two are outlined in blue, using dlx2a expression as a guide. Markers are indicated on the left panel of each row, and treatments indicated above each column. Anterior is leftwards, dorsal upwards. (O) In hand2S6, ectopic eng2 is indicated with an arrow. (U) Volumes (y-axis) of dlx2a, dlx4a and eng2 expression. Error bars are 95% confidence intervals, from ANOVA. Measurements were made on confocal stacks of randomly selected fish, using the `find objects by intensity′ function in Volocity software. Intensity thresholds were adjusted from fish to fish, in order to accurately identify expression. There are no significant differences between fish classes in average intensity levels. Each bar shows the combined volumes of arches one to three, because these arches were sometimes identified as one object by the software. Scale bar: 100 μm.
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