DEV02439 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. zmsn1 is required for epiboly as well as convergent extension. (A) RT-PCR analysis of msn1 mRNA of embryos injected at the one-cell stage with msn1MO-splice (MO). Primers (P1/P2) flanking the region result in a single 400 bp band in the case of control embryos. In the case of morphants, the level of this band is strongly reduced and a second band (red arrowhead) is visible. The second band results from the use of an alternative splice donor. (B-E) mRNA expression patterns of bmp-4 (B,C, dorsal is to the right) and gsc (D, E, view onto the dorsal side) to determine dorsal-ventral patterning. (F-K) Analysis of the degree of epiboly and convergent extension. Bright-field image of a control embryo at bud stage (F) and an embryo injected with msn1MO-splice at the one cell stage (G). Simultaneous analysis of the mRNA patterns of the axial markers dlx3, hgg1 and ntl in a control (H,J) and morphant (I,K) embryo. (L) Quantification of the penetrance of gastrulaltion defects in uninjected control embryos (co), embryos injected with msn1MO-splice (MO), and embryos injected with msn1MO-splice-5bp (MOcon). Shown are the percentages of embryos displaying delayed epiboly (80-95% epiboly) when uninjected embryos had reached the 100% epiboly stage Scale bar: 100 μm.

• Movie 1 -

  Movie 1. Time-lapse movie of a zebrafish control embryo. A wild-type embryo expressing membrane-GFP and cytosolic GFP in all cells was imaged on a multi-photon confocal microscope (see Materials and methods). Alexa 488 Histone H1 had been injected into the YSL to label YSL nuclei. A single focal plane is shown, allowing the observation of the EVL cells and some deep cells. Individual images from this movie are shown in Fig. 4. Duration: 78 minutes.

• Movie 2 -

  Movie 2. Time-lapse movie of a YSL-injected msn1MO embryo. A wild-type embryo expressing membrane-GFP and cytosolic GFP and co-injected into the YSL with msn1MO and Alexa 488 Histone H1. A projection of multiple focal planes is shown. Images from individual focal planes of this movie are shown in Fig. 4. Duration: 132 minutes.

• Movie 3 -

  Movie 3. Time-lapse movie of a Drosophila control embryo during dorsal closure. A control embryo expressing GFP-Actin in the engrailed stripes was imaged on a confocal microscope during the final phase of dorsal closure (see Materials and methods). A dorsal view of the embryo is shown with anterior to the left. Duration: 38 minutes.

• Movie 4 -

  Movie 4. Time-lapse movie of a msn¹⁷² Drosophila embryo during dorsal closure. Confocal movie of an embryo homozygous for msn¹⁷² and expressing GFP-Actin in the engrailed stripes. The field of view was shifted during the movie to capture a larger area of the embryo. Duration: 101 minutes.