Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie01 - Movie 1. The localization of Msps-GFP is spatially regulated in interphase Drosophila S2 cells. S2 cell transfected with full length Msps-GFP (left) and mCherry-�-tubulin (right). Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie02 - Movie 2. Msps-GFP exhibits dynamic movements along the lattice of peripheral microtubules. S2 cell transfected with Msps-GFP (left) and mCherry-Tubulin (right) showing the dynamic movement of discontinuous GFP-punctae on peripheral microtubules. Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie03 - Movie 3. Msps-GFP associates with growing and shrinking microtubules. S2 cell transfected with Msps-GFP (left) and mCherry-Tubulin (right) shows growing and shrinking microtubule association. Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie04 - Movie 4. Msps localizes to plus ends and microtubule lattice, but not centrioles, in an EB1-dependent manner. S2 cell transfected with Msps-GFP and treated with either control (left) or EB1 dsRNA (right). Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie05 - Movie 5. Msps TOG domains are sufficient to partially rescue microtubule polymerization and EB1 dynamics. S2 cells transfect with EB1::EB1-GFP and a COOH-terminally tagged Msps construct (inset, bottom left), were treated with control or Msps dsRNA. Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie06 - Movie 6. Msps TOG domains 1-2 and 3-4 provide little rescue of microtubule polymerization and EB1 dynamics. S2 cells transfect with EB1::EB1-GFP and a COOH-terminally tagged Msps construct (inset, bottom left), were treated with control or Msps dsRNA. Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie07 - Movie 7. Double Mut Msps-GFP does not associate with the lattice of peripheral microtubules and results in gross alterations to the microtubule cytoskeleton. S2 cells treated with Msps 5�UTR dsRNA for 7 days and transfected with either wildtype Msps-GFP (top, left) or Double Mut Msps-GFP (bottom, left) and mCherry-Tubulin (right). Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).
- Movie08 - Movie 8. Mutation in the linker2 and linker4 motifs of full length Msps affect peripheral lattice association. S2 cell transfected with either wildtype full-length Msps-GFP (left, green) or with Double Mutant full-length Msps-GFP (Double Mut Msps-GFP) and mCherry-Tubulin (right, red). Images were acquired at three second intervals using spinning-disc confocal microscopy (Yokogawa, Perkin Elmer).