DEV02417 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. ICD and ESR6e repress radial intercalation. mRFP-labeled outer cells were transplanted at stage 10 onto the inner layer of host embryos expressing ICD (B,E), or ESR6e (C) along with mGFP, or with just mGFP (A,D) as a control. At either stage 28 (A-C, tadpole) or stage 20 (D-E, neurula), the embryos were fixed, stained with antibody to ciliated cells (CC, blue) and imaged by confocal microscopy. ESR6e strongly inhibits the formation of ciliated cells, and represses intercalation of INCs (a few remaining INCs are shown in C), as quantified in F. ICD eliminates ciliated cells (B,F), but not INCs at this stage. However, many of the INCs detected at stage 28 in ICD-injected embryos have a small apical domain, suggesting that their intercalation was delayed. In line with this idea, ICD strongly inhibits intercalation when scored at stage 20 (compare E with D). These data indicate that activation of Notch signaling and, thus ESR6e expression both represses ciliated cell differentiation and delays the intercalation of non-ciliated cells.

• Movie 1 -

  Movie. 1. Morphology of CCs and INCs. mRFP-labeled outer cells were transplanted at stage 10 onto the inner layer of host embryos expressing mGFP. At tailbud stages, transplants were imaged live using a confocal microscope. Shown is a 3D reconstruction of a CC (left panel) and an INC (right panel) based on series of maximal projections through a z-stack of images collected at 0.5 μm intervals. Scale bar: 10 μm