| |
Fig. 4. Hairy2 affects the expression of neural border genes. Hairy2-GR mRNA overexpression (500 pg) expands Msx1 (b, 64% n = 22), Pax3 (d, 62% n = 17) and strongly increases Zic1 (f, g, 78% n = 14), SoxD (i, j, 100% n = 11). Injection of Hairy2 MOs (10 ng) reduces Msx1 (a, 60% n = 26) and expands Pax3 (c, 58% n = 31) and Zic1 (e, 53% n = 17). SoxD appears unaffected (h, 80% normal n = 20). Embryos are shown in dorsal (a–f, h, i) or lateral view (g, j), with inset showing control sides. β-galactosidase was used as a lineage tracer. |
| |
Fig. 7. Id3 is required for Hairy2 to promote NC formation. (A) Hairy2-GR (500 pg) injection induces Sox10 (a; 55% increased, n = 24) in tailbuds and upregulates SoxD (e, 88%, n = 16), Pax3 (h, 65%, n = 32), Msx1 (k, 59%, n = 27) and Zic1 (n, 63%, n = 32) in neurula embryos. Hairy2-GR and Id3 MO co-injection(15 ng) decreases Sox10 (60% inhibition, n = 27) in embryos but still induces SoxD (g, 70% increased n = 20), Pax3 (j, 62%, n = 29), Msx1 (m, 49, n = 35) and Zic1(p, 67%, n = 33). Control minus DEX shows that the Id3 MO efficiently inhibits Sox10 (50% inhibited, n = 15;) and has no effect on SoxD (f, 76% normal n = 17) Pax3 (i, 70%, n = 23), Msx1 (l, 85%, n = 21) and Zic1(o, 65%, n = 23). Hairy2-GR was also unable to induce Sox10 with Id3 MO in NC induced animal explants (d). (B) Id3-GR (1 ng) partially rescues Snail2 and Sox10 in Hairy2 (10 ng) depleted embryos (a, b, d, e) and in NC induced animal caps explants analysed by Q-PCR (c, f). Respective inhibition and rescue: (a) 68% inhibited, n = 26; (b) 89% rescued, n = 27; (d) 56% inhibited, n = 18; (e) 75% rescued; n = 30. Dorso-anterior views of embryos with injected side to the right (a, b) or lateral views (d, e) of embryos are shown. β-galactosidase was used as a lineage tracer. |
