JCS018937 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1: Alignment of Xenopus NEDD1, human NEDD1 and Dgp71WD. Amino acid residue 300 (which constitutes the end of XNEDD1-N and the beginning of XNEDD1-C) is indicated by ‘300’. Ser418, which is phosphorylated in human NEDD1 (Lüders et al., 2005), and the corresponding serine residue in Xenopus NEDD1 are indicated by a red box.

• Supplemental Figure S2 -

  Fig. S2: Localization throughout the cell cycle of GFP-XNEDD1 expressed in Xenopus tissue culture cells. Cell cycle stages are indicated on the left. Scale bar, 10 µm.

• Supplemental Figure S3 -

  Fig. S3: Several examples of Xenopus NEDD1 (XNEDD1; green) and γ-tubuln (red) double staining at sperm tips (blue). XNEDD1 generally stains two spots, whereas γ-tubulin stains a larger area. These sperm tips were not incubated in egg extract. Scale bar, 2 µm.

• Supplemental Figure S4 -

  Fig. S4: Recombinant Xenopus NEDD1 (XNEDD1) localizes to microtubules like endogenous XNEDD1. XNEDD1-depleted extract was resonstituted with buffer, full-length XNEDD, or XNEDD-C as indicated on the left. Samples were spun onto coverslips and processed for immunofluorescence using antibodies against XNEDD, α-tubulin or DNA, as indicated. XNEDD is green in the overlays, tubulin is red, and DNA is blue. Scale bar, 10 µm.

• Supplemental Figure S5 -

  Fig. S5: Xenopus NEDD1 (XNEDD1) depletion disrupts taxol-induced microtubule aster assembly. (A) XNEDD1 localizes to the center of taxol-induced asters. Scale bar, 10 µm. (B) Taxol-induced microtubule structures assembled in mock-depleted (mock-Δ) or XNEDD1-depleted (XNEDD1-Δ) extracts. Scale bar, 10 µm.