DEV01669 Supplementary Material

Files in this Data Supplement:

• Movie 1 -

  Movie 1. Cellularisation of wild-type embryo observed with DIC optics.

• Movie 2 -

  Movie 2. Cellularisation of embryo from RhoGEF2^1.1 germline clones observed with DIC optics.

• Movie 3 -

  Movie 3. Cellularisation of embryos from dia germline clones observed with DIC optics.

• Supplemental Figure S1 -

  Fig. S1. Basal closure in embryos from RhoGEF2 and dia germline clones. (A) Wild-type embryo and embryos from (B) RhoGEF^1.1 and (C) dia germline clones in later stage of cellularisation stained for Slam protein (green, white) and DNA (blue) to visualise the widened base of the furrow canal. Scale bar: 10 mm.

• Supplemental Figure S2 -

  Fig. S2. Phenocopies of sry-a and nullo by dsRNA injection. (A) Embryo from RhoGEF2 germline clones injected with control dsRNA (Bsg25D). Wild-type embryos injected with (B) sry-a and (C) nullo dsRNA. Embryos were fixed and stained with phalloidin (green) and DNA (blue). Scale bar: 10 mm.

• Supplemental Figure S3 -

  Fig. S3. Myosin II localisation depends on RhoGEF2 and dia. (A,C) MyoII localises to the furrow canal in wild-type embryos. MyoII levels are reduced in embryos from (B) dia and (D) RhoGEF2⁰⁴²⁹¹ germline clones. MyoII is more strongly reduced in dia mutants than RhoGEF2 mutants but recovered to almost wild-type levels during the second half of cellularisation. Embryos of the indicated genotype were stained for MyoII (green) and DNA (blue). Wild-type and mutant embryos were processed in one tube and identified after mounting.

• Supplemental Figure S4 -

  Fig. S4. RhoGEF2 and Nullo protein localisation are independent of each other. Sry-a localisation is similar in (A) wild-type embryos and in embryos from (B) RhoGEF2^1.1 and (C) dia germline clones. RhoGEF2 protein and Dia protein are concentrated at the furrow canal in (D,F) nullo embryos and RhoGEF2, and in (E) embryos from dia germline clones. Embryos of indicated genotypes were stained for the indicated proteins (Sry-a, red; RhoGEF2, green; Dia, yellow and white in right half of the panels; DNA, blue).

• Supplemental Figure S5 -

  Fig. S5. Purified recombinant proteins for biochemical analysis. (A,B) GST or ZZ fusion proteins (1 mg) of the indicated proteins analysed by SDS PAGE and stained with Coomassie dye.

• Supplemental Figure S6 -

  Fig. S6. FH domins of Dia induce actin polymerisation. Kinetics of actin (3 mM) polymerisation induced by increasing concentrations (20 nM, 100 nM, 1 mM) of ZZ-DN519.