Free microtubule sliding in cnm67delta mutant cells expressing GFP-Tubulin (GFP-TUB1; left panel, green in right panel merge) and RFP-Spc97 (a component of the gamma-tubulin complex, marks the minus end; middle panel, red in right panel merge). Movie plays at 50x real time and is movie S9 in Proc Natl Acad Sci. 2009. 106: 5147-5152.
S. cerevisiae (MATa SPC97-tdimer2::KanR cnm67delta::TRP1 ura3-52 lys2-801 leu2-delta1::GFP-TUB1::LEU2 his3-delta200 trp1-delta63) were grown to midlog phase and mounted on agarose pads for microscopy. Images were collected on an Olympus IX70 inverted fluorescence microscope with a 100 N.A. 1.35 oil objective lens and a CoolSNAP HQ camera (Roper Scientific), using QED software (Media Cybernetics). Timelapse images were captured at 5-s intervals.
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