DEV01843 Supplementary Material

Files in this Data Supplement:

• Supplemental Tables (Adobe PDF)
• Supplemental Figure 1 -

  Fig. S1. Anillin localizes to the stalks and rings that connect newly cellularized cells to the yolk mass. Scale bars: 10 mm in A; 5 mm in B,C. (A) A methanol-fixed gastrulating embryo, sectioned perpendicular to the embryo surface and probed with antibody against Anillin. The asterisk indicates the cephalic furrow and the small arrowhead to its left indicates stalks of cells at the edge of the furrow. The large arrowhead (bottom center) indicates stalks from cells of the ventral furrow. (B,C) High-magnification images of stalks imaged on two different planes (same embryo as in A). (B) Cell stalks. Several stalks appear connected together via a tube-like structure that contains Anillin (arrowheads). (C) Images in a plane above the stalks (more toward the cell base) show small contracted rings that are rich in Anillin.

• Supplemental Figure 2 -

  Fig. S2. Anillin and Peanut localization in a weak anillin allele. Indirect immunfluorescence of cold methanol fixed embryos using laser confocal imaging. Scale bar is 5 mm. (A) anillin^RV/RV-derived embryo in late cellularization, sectioned normal to embryo surface (A) and sectioned parallel to the surface at the cellularization front (a,a¢,a¢¢). In contrast to the strong alleles (Fig. 4), Anillin does colocalize with Peanut, and contractile rings form (a,a¢,a¢¢). Ectopic puncta of Peanut are observed above the furrow front (A, arrowheads) and between contractile rings (a¢¢, arrowhead). In some contractile rings, Anillin and Peanut staining is weak (a,a¢,a¢¢, white asterisk). (B) anillin^RV/RV-derived embryo during gastrulation sectioned parallel to the surface at the cellularization front (B,b,b¢¢). Rings contain both Anillin and Peanut, often not colocalized (b¢). Septin rings can exist independently of Anillin (B,b, b¢, arrowheads). Some areas lack stalk/ring structures (B,b,b¢, black asterisk). Rings are comparable in size with septin rings observed by Kinoshita et al. (Kinoshita et al., 2002) in mammalian cells and in vitro.

• Supplemental Figure 3 -

  Fig. S3. Neurotactin (integral membrane protein) localization during cellularization. Heat and methanol fixed embryos. All images are sections normal to the embryo surface. Scale bar: 5 mm in A-D; 15 mm in c and d. (A) Wild-type embryo in early fast phase. Black arrowhead indicates a furrow canal. White arrowhead indicates a solid line of Neurotactin localizing at the lateral membrane. (B) anillin^PQ/RS-derived embryo in early fast phase. Neurotactin appears punctate and fuzzy (arrowhead), indicating problems with membrane stability or insertion. Furrow canals are not evident. (C) Wild-type embryo in late cellularization. Black arrowhead indicates a furrow canal. Boxed area is shown in higher magnification on the right (c). White arrowhead in c indicates solid line of Neurotactin localized at the lateral membrane. (D) anillin^PQ/RS-derived embryo in late cellularization, furrow canals are not evident. White asterisk indicates nucleus moving to the embryo interior. Boxed area is shown at high magnification on the right (d). Neurotactin appears punctate and fuzzy (arrowhead in d), indicating problems with membrane stability or insertion.