JCS038497 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. TLNcdVenus in the mitotic spindle. TLncdVenus; ncdmRFP ca^nd cycle 9 embryo showing faint TLNcdVenus spindle fluorescence and little or no TLNcdVenus centrosome fluorescence (arrows). Scale bar: 10 µm.

• Supplemental Figure S2 -

  Fig. S2. FLNcdVenus and HLNcdVenus fluorescence recovery data fit to different models. (A) FLNcdVenus small (w=1.3 µm) and large (w=2.66 µm) ROI FRAP data at the spindle equator (magenta) fit concurrently to a two-state binding model (bright green) (Sprague et al., 2004), compared with the new diffusion-binding-transport model (dark green). (B) HLNcdVenus small (w=1.3 µm) and large (w=2.66 µm) ROI FRAP data at the spindle equator (purple) fit concurrently to a two-state binding model (bright green) or (C) a diffusion-binding model (cyan) (Sprague et al., 2004), compared with the new diffusion-binding-transport model (dark green).

• Movie 1 -

  Movie 1. HLNcdVenus in mitosis. HLncdVenus cycle 10 cleavage division from prometaphase to telophase. Images were recorded at 30 second intervals and play at 10 frames per second. Scale bar: 10 µm.

• Movie 2 -

  Movie 2. TLNcdVenus in mitosis. TLncdVenus cycle 9 cleavage division from prometaphase through telophase. Images were recorded at 15 second intervals and play at 20 frames per second. Scale bar: 10 µm.

• Movie 3 -

  Movie 3. HLNcdVenus mitotic spindle FRAP assay with a small ROI (w=1.3 µm). Prebleach images (6) are followed by 494 recovery images (see Fig. 3C,D) after bleaching. Images were recorded at ∼165 mseconds/frame and play at 20 frames per second. Scale bar: 3 µm.

• Movie 4 -

  Movie 4. HLNcdVenus mitotic spindle FRAP assay with a large ROI (w=2.66 µm). Prebleach images (6) are followed by 494 recovery images (see Fig. 3C,D) after bleaching. Images were recorded at ∼165 mseconds/frame and play at 20 frames per second. Scale bar: 3 µm.

• Movie 5 -

  Movie 5. Ncd transport towards the equator in mitosis. FLncdVenus mitotic spindle from early metaphase through anaphase cropped to delete centrosomes, astral microtubules, and cytoplasm. Images were acquired at 2.32 second intervals and play at 20 frames per second. Scale bar: 3 µm.