DEV000802 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Nucleotide alignment covering the first thousand bases of the spliced K09H11.3 and Y75B7AL.4 genes. This region covers in both cases the predicted RhoGAP domain (approximately bp 141-831 in the K09H11.3 sequence). Clearly visible is the high similarity between both sequences. The alignment was done by using ClustalW.

• Supplemental Figure S2 -

  Fig. S2. Mitosis and nuclear division is fine in rga-3/4(RNAi) and cytokinesis is sometimes affected. (A) Wild-type and rga-3/4(RNAi) embryos expressing H2B::GFP were stained with FM4-64 to visualize membranes and nuclei at the same time. rga-3/4(RNAi) embryos showed cytokinesis defects and formation of cytoplasts (white arrows). (B) Nuclear division is not affected in rga-3/4(RNAi) embryos. Chromatin is visualized by H2B::GFP. (C) The formation of the mitotic spindle is not affected in rga-3/4(RNAi) embryos. The mitotic spindle was visualized using a strain expressing beta-tubulin::GFP, although spindle rotation is sometimes affected (beta-tubulin::GFP and H2B::GFP).

• Supplemental Figure S3 -

  Fig. S3. Analysis of GFP::RGA-3-expressing worms. (A) gonad migration defect in L4 larvae. The black arrows point to the two different gonad arms. (B) Gonad migration in adults. One gonad arm overshoots and bends into the uterus. The other gonad arm has an unusual swelling in the tip area. The white arrows point to the same gonad arm in two different frames. The black arrow points to the swelling in the tip of the second gonad arm. (C) GFP.:RGA-3 expression in the gonad and oocytes. The upper panel shows the gonad of a larva. The gonad seems crippled. The lower panel displays a part of an adult worm. The strong fluorescence is derived from auto-fluorescing gut granules. The arrowheads point to two oocytes positive for GFP::RGA-3.

• Movie 1 -

  Movie 1. Wild-type fertilized zygote up to pro-nuclear meeting.

• Movie 2 -

  Movie 2. Knockdown of RGA-3/4 in the one-cell embryo.

• Movie 3 -

  Movie 3. rho-1(RNAi) phenotype in the one-cell embryo.

• Movie 4 -

  Movie 4. Phenotype in the early embryo of rho-1(RNAi) rga-3/4(RNAi).

• Movie 5 -

  Movie 5. cdc-42(RNAi) phenotype in the one-cell embryo.

• Movie 6 -

  Movie 6. Phenotype in the early embryo of cdc42(RNAi) rga-3/4(RNAi).

• Movie 7 -

  Movie 7. let-502^−/− phenotype in the one-cell embryo.

• Movie 8 -

  Movie 8. Phenotype in the early embryo of let-502^−/− rga-3/4(RNAi).

• Movie 9 -

  Movie 9. GFP::PAR-6 localization in an one-cell rga-3/4(RNAi) embryo.

• Movie 10 -

  Movie 10. GFP::PAR-6 localization in an one-cell cyk-4(RNAi) embryo.