DEV001602 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Proliferation of EGFP⁺ cells in mib^−/−;Tg(olig2:egfp) embryos. Dorsal views of the posterior neural plate at 11 (A,B), 12 (C,D) and 14 hpf (E,F) and lateral views of spinal cord at 18 hpf (G,H) in Tg(olig2:egfp) (A,C,E,G) and mib^−/−;Tg(olig2:egfp) embryos (B,D,F,H). Green, blue and red channels mark EGFP⁺ cells, PH3⁺ mitotic cells and Hu⁺ postmitotic neurons, respectively. Arrows indicate PH3⁺ EGFP⁺ cells that underwent M phase of cell division. (I) Average of PH3⁺ EGFP⁺ cells in control (squares) and mib^−/− embryos (triangles) (n=4 animals each). Error bars represent s.e.m. Scale bar: 50 μm

• Supplemental Figure S2 -

  Fig. S2. Recovery of Notch signaling after transient inhibition. Dorsal views of flat-mounted zebrafish embryos, labeled by her4. A′-I′ are magnified views of boxed areas in A-I. (A,A′) Control embryo showing normal expression of her4 at 12 hpf. Bracket marks her4-expressing neuroectoderm cells. (B,B′) Continuous DAPT incubation caused significant reduction in her4 expression in posterior neuroectoderm (bracket). (C,C′) Embryos incubated with DAPT from 6 to 11 hpf re-expressed her4 (bracket) at 12 hpf. (D-F′) 11 hpf embryos. (D,D′) Control embryo. (E,E′) After heat shock at 9.5 hpf, Tg(hsp70l:XdnSu(H)myc) embryos did not express her4. (F,F′) After heat shock at 6 hpf, transgenic embryos did not express her4. (G-I′) 12 hpf embryos. (G,G′) Control embryo. (H,H′) After heat shock at 9.5 hpf, transgenic embryos did not express her4. (I,I′) After heat shock at 6 hpf, her4 expression was evident in posterior neuroectoderm in transgenic embryos. Scale bar: in A, 200 μm for A-I; in A′, 100 μm for A′-I′.