Figure 6:

Increasing XB5850668.L has minimal effects on ectodermal gene expression domains.

A: The expression domains of neural border genes were mostly unaffected by increased XB5850668.L. However, a small percentage of those associated later in development with the lateral neural plate and neural crest (pax3, msx1, anterior views) showed reduced intensity of expression on the mRNA-injected side of the embryo (pink lineage-labeled nuclei) compared to the control side (ctrl) of the same embryo. dlx5, which later in development is associated with epidermis and PPE, was unaffected by increased XB5850668.L (anterior view); the same was found for tfap2a expression (shown in 2B). Dorsal is to the top of each image.

B: Percentage of embryos in which neural border expression domains were reduced (dark blue), broader (medium blue), or did not change (light blue) when analyzed at the end of gastrulation (st 13). The number of embryos analyzed for each gene at the top of each bar.

C: The expression domains of neural plate genes were mostly unaffected by increased XB5850668.L. However, a small percentage of embryos showed broader neural plate domains on the mRNA-injected side of the embryo (pink lineage-labeled nuclei) compared to the control side (ctrl) of the same embryo. Arrows in sox11 show its PPE expression domain on the control (black) and mRNA-injected (red) sides. In this case, increased XB5850668.L resulted in a narrower sox11 PPE domain. Anterior views with dorsal to the top.

D: Percentage of embryos in which neural plate expression domains were reduced (dark blue), broader (medium blue) or did not change (light blue) when analyzed at neural plate stages (st 16-18). The number of embryos analyzed for each gene is at the top of each bar.

E: The expression of epidermis-specific keratin (krt12.4) was reduced on the mRNA-injected side in only 25% of embryos; there was no change in expression in 75% of embryos (n=56). Anterior view with dorsal to the top.

F: The expression domains of neural crest genes were mostly unaffected by increased XB5850668.L (see 6H for sox9). However, small percentages of embryos showed a broader domain (pax3, foxd3) or reduced staining intensity (sox9) on the mRNA-injected side of the embryo (pink lineage-labeled nuclei) compared to the control side (ctrl) of the same embryo. For pax3, both the neural crest (bracket) and hatching gland (arrow) domains were affected. Arrows in sox9 show its otic placode domain on the control (black) and mRNA-injected (red) sides, which frequently was reduced (see also 6H, I). Anterior views with dorsal to the top.

G: Percentage of embryos in which neural crest expression domains were reduced (dark blue), broader (medium blue) or did not change (light blue) when analyzed at neural plate stages (st 16-18). The number of embryos analyzed for each gene is at the top of each bar.

H: The expression domains of PPE/placode genes were mostly unaffected by increased XB5850668.L. However, small percentages of embryos showed a broader domain (sox11) or a reduced domain (sox11 in 6C; sox9 and also 6F) on the mRNA-injected side of the embryo (pink lineage-labeled nuclei) compared to the control side (ctrl) of the same embryo. Arrows indicate PPE/placode domains on the control (black) and mRNA-injected (red) sides. Anterior views with dorsal to the top.

I: Percentage of embryos in which PPE expression domains were reduced (dark blue), broader (medium blue) or did not change (light blue) when analyzed at neural plate stages (st 16-18). The number of embryos analyzed for each gene is at the top of each bar.