Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie 8 - Video – Cells expressing the YNIL mutant form of eGFP‐Tm1J exhibit spindle morphology defects and delayed anaphase progression. S2 cells stably expressing the YNIL mutant version of eGFP‐Tm1J (green) and mCherry‐Tubulin (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by visualizing the mitotic spindle using mCherry‐Tubulin.
- Movie 3 - Video – Tm1A and Myosin II co‐localize to retracting membrane blebs. S2 cells stably expressing myosin‐eGFP (green) and mCherry‐Tm1A (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Blebbing cells were identified by cell morphology.
- Movie 10 - Video – Tm1J localizes to the centrosomes, kinetochores, central spindle and midpoint during mitosis and returns to cytoplasmic rings at Golgi after cell division. S2 cells stably expressing eGFP‐Tm1J (green) and mCherry‐Tubulin (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by visualizing the mitotic spindle using mCherry‐Tubulin.
- Movie 1 - Video – Golgi‐associated Tm1J rings disassemble during prophase and fully disperse after nuclear envelope breakdown. S2 cells stably expressing eGFP‐Tm1J (green) and mCherry‐Tubulin (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy.
- Movie 9 - Video – Fringe‐eGFP containing Golgi stacks disassemble after nuclear envelope breakdown (NEBD). S2 cells stably expressing Fringe‐eGFP (green) and mCherry‐Tubulin (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy.
- Movie 7 - Video – Tm1J still localizes to kinetochores but does not localize to the central spindle or midpoint in the absence of Diaphanous. Diaphanous was depleted from S2 cells stably expressing eGFP‐Tm1J (green) and mCherry‐Tubulin (red) using RNAi. Cells were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by cell morphology and mCherry‐Tubulin fluorescence.
- Movie 6 - Video – Tm1A initially localizes to the equatorial cortex but does not remain there in the absence of Diaphanous. Diaphanous was depleted from S2 cells stably expressing mCherry‐Tm1A (shown) and eGFP‐Tm1J (not shown) using RNAi. Cells were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by cell morphology and eGFP‐Tm1J fluorescence.
- Movie 5 - Video – Tm1A and Diaphanous only partially overlap at the equator of dividing cells. S2 cells stably expressing Diaphanous‐eGFP (green) and mCherry‐Tm1A (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by cell morphology.
- Movie 4 - Video – Tm1A and Myosin II co‐localize at the equatorial cortex in dividing cells. S2 cells stably expressing myosin‐eGFP (green) and mCherry‐Tm1A (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy. Dividing cells were identified by cell morphology.
- Movie 2 - Video – Golgi‐associated Diaphanous rings disassemble during prophase and fully disperse after nuclear envelope breakdown. S2 cells stably expressing Dia‐eGFP (green) and mCherry‐Tubulin (red) were plated onto poly‐D‐lysine coated coverslips and imaged by time‐lapse confocal microscopy.