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. 2016 May 1;143(9):1523–1533. doi: 10.1242/dev.128165

Fig. 4.

Fig. 4.

sFRP1/5 and Dkk3 size the sea urchin ANE. (A) foxq2, nkx3.2 and six3 expression at mesenchyme blastula stages in control (a-c), sFRP1/5 morpholino knockdown (e-g) and Dkk3 morpholino knockdown (i-k) embryos. (d,h,l) Serotonergic neurons (green) in control, sFRP1/5 and Dkk3 knockdown 80 hpf pluteus larvae. The white line indicates the territory occupied by the serotonergic neurons (arrowheads). (B) The angle α shown in A was measured for the most posterior boundaries of the selected ANE territory markers for ∼24 embryos from three batches using ImageJ. Solid lines indicate the posterior boundaries of each ANE marker used to measure α; the dashed lines show the most anterior boundary of six3 expression. Volume=0.5(1−cos α/2) was used to calculate the percentage of the surface area (±s.e.m.) occupied by the inner (foxq2, nkx3.2) and outer (six3) ANE territories in control, sFRP1/5 and Dkk3 morphants. (C) qPCR analysis showing that the expression of ANE regulatory genes is upregulated in sFRP1/5 and Dkk3 morphants. The y-axis shows the fold change in gene expression level in sFRP1/5 and Dkk3 morphants. The bars represent data using two different morpholinos. MO, morpholino. Scale bars: 20 µm.