Free microtubule sliding in nip100deltaCAP-Gly cnm67delta double mutant cells. Movie plays at 50x real time and is movie S12. Compare with movie S11 (CIL# 23601). Data analysis for movies S11 and S12 in Figure S4B of Proc Natl Acad Sci. 2009. 106: 5147-5152.
S. cerevisiae (MATalpha NIP100deltaCAP-Gly-13myc::KanMX6 cnm67delta::TRP1 ura3-52 lys2-801 leu2-delta1::GFP-TUB1::LEU2 his3-delta200 trp1-delta63) expressing GFP-tubulin (GFP-TUB1) were grown to midlog phase and mounted on agarose pads for microscopy. Images were captured on an Olympus Bmax-60F microscope equipped with a 1.35NA 100× UPlanApo objective, spinning disc Confocal Scanner Unit (CSU10), Picarro Cyan laser (488 nm), and a Stanford Photonics XR-Mega10 ICCD camera, by using QED software (Media Cybernetics). Image analysis was performed by using ImageJ. Timelapse images were captured at 10-s intervals for 15 min and each image in the movie represents a composite of 9 planes separated by 500 nm.
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