DEV027938 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig S1. Comparison of the expression of deltaD transcript following loss of Notch signalling. (A-F′) A comparison of deltaD transcripts in wild-type sibling zebrafish embryos (A,A’) with that in mib mutants (B,B’), in un-injected embryos (C,C’) with Notch MO morphants (D,D’), and in DMSO-treated controls (E,E’) with DAPT treated embryos (F,F’) at the tail bud stage. Accompanying panels on the right (A′-F′) are magnified images of neurogenic domains in hindbrain delineated with rectangles in A-F. deltaD transcripts were not significantly increased at the tail bud stage following reduced Notch signaling.

• Supplemental Figure S2 -

  Fig S2. Notch interaction regulates DeltaA endocytosis. (A,B) Comparison of zdD2 detection with a fluorescent secondary with standard gain used for most cases in this study (A), and with higher gain (B). (C-F′) zdD2 distribution visualized using higher gain in aei^AG49 homozygous embryos (C,E,E’) and in aei^AG49 homozygous embryos co-injected with notch1a and notch3 (D,F,F’). There remains faint zdD2 staining in aei^AG49 homozygous embryos (C), which is easier to see when notch1a and notch3 are knocked down (D). (E-F′) aei^AG49 homozygous embryo with (F,F′) or without (E,E′) notch morpholinos are double stained with zdD2 (grey in E,F, green in E′,F′) and plasma membrane marker β-catenin (red in E’,F’). Magnified images of two longitudinal neurogenic domains in the prospective hindbrain. The setting with high gain was used to take these images. With standard gain there is almost no signal detected in aei^AG49 homozygous embryos. DeltaA remains at the plasma membrane in Notch MO (F,F′).