Quantifying Microtubule Assembly Dynamics

Cells were transfected with EB3-GFP and RFP-CenpB to label assembling MTs and centromeres, respectively (25 (link)). Movies of MT assembly were collected over 30 sec at 500 ms intervals 10 min after drug application (0.5 µM ST-11, 3 µM ST-11, or 1 µM nocodazole) on a Deltavision microscope system (Applied Precision, Issaquah, WA). Images were deconvolved using SoftWorx 5.0 (Applied Precision). MT assembly rates were scored in interphase cells using Fiji TrackMate (26 (link)).

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Cherry A.E., Haas B.R., Naydenov A.V., Fung S., Xu C., Swinney K., Wagenbach M., Freeling J., Canton D.A., Coy J., Horne E.A., Rickman B., Vicente J.J., Scott J.D., Ho R.J., Liggitt D., Wordeman L, & Stella N. (2016). ST-11: a new brain-penetrant microtubule-destabilizing agent with therapeutic potential for glioblastoma multiforme. Molecular cancer therapeutics, 15(9), 2018-2029.

Publication 2016

Deltavision microscope system SoftWorx 5.0

Cells Cenpb Centromeres Drug Interphase Microscope Nocodazole

Corresponding Organization : University of Washington

Other organizations : Howard Hughes Medical Institute

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Variable analysis

independent variables

Drug application (0.5 µM ST-11, 3 µM ST-11, or 1 µM nocodazole)

dependent variables

MT assembly rates

control variables

Cells were transfected with EB3-GFP and RFP-CenpB to label assembling MTs and centromeres, respectively
Movies of MT assembly were collected over 30 sec at 500 ms intervals 10 min after drug application
Images were deconvolved using SoftWorx 5.0 (Applied Precision)
MT assembly rates were scored in interphase cells using Fiji TrackMate

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