Shroom family proteins regulate -tubulin distribution and microtubule architecture during epithelial cell shape change

DEV02828 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Patterns of apical constriction in the Xenopus neural plate. The highly constricted cells of the hingepoints are strictly ordered, and the constricting cells form anteroposterior arrays. This is most easily seen by comparing surface views (A,a′) with cross-sectional views (B,b′). In a′ and b′, hingepoint cells are indicated in red and intermediate cells in blue; the midline is indicated in purple.

• Supplemental Figure 2 -

  Fig. S2. Shroom3 is necessary for the proper localization of γ-tubulin in neuroepithelial cells. (A) Cross-section through the closing anterior neural tube of a stage 16 embryo injected unilaterally (right) with Shroom3-MO. Embryo has been stained for both γ-tubulin (A) and α-tubulin (a′). Merged image is shown in panel a′′; green, γ-tubulin and red, α-tubulin. (B) Cross-section through the closing anterior neural tube of a stage 18 embryo injected on one side (right) with Shroom3-MO. (C) Cross-section through the closing anterior neural tube of a stage 18 embryo injected on one side (right) with mRNA encoding Shrm754-1108.

• Supplemental Figure 3 -

  Fig. S3. Western blotting of γ-tubulin levels in manipulated embryos. (A) Western blot for γ-tubulin in isolated dorsal tissues (notochord and neural tube) of stage 16 embryos injected bilaterally with Shroom3-MO. (B) Western blot for γ-tubulin in isolated stage eight animal caps from embryos injected with Shroom3 mRNA into all four blastomeres at the four-cell stage.

• Supplemental Figure 4 -

  Fig. S4. Xenopus Shroom3. (A) Shroom3 encodes a protein with high similarity to mouse Shroom3S isoform. (B) Control embryo showing surface view of the superficial blastomeres epithelium; (b′) phalloidin staining reveals only junctional actin. Expression of mShroom3L (C), mShroom3S (D), or XShroom3 (E) is sufficient to induce both apical constriction and apical actin accumulation (c′,d′,e′).