Journal: Molecular Biology of the Cell
Article Title: Spatial regulation of astral microtubule dynamics by Kif18B in PtK cells
doi: 10.1091/mbc.E16-04-0254
Figure Lengend Snippet: Kif18B knockdown causes an increase in the lifetime of astral MTs and a decrease in MT growth rate in general. (A, B) First frame of EB1-GFP image series of spindles in PtK1 cells transfected with (A) control or (B) Kif18B-2 siRNA. Scale bar, 10 μm. (C, D) The plusTipTracker software was used to detect and link EB1 comets from time-lapse images of spindles in A and B. Tracked comets were divided into three subspindle regions and color coded accordingly: astral MTs to the left and right of spindle poles (aster_lr) are in magenta, astral MTs in the top and bottom region of the spindle body (aster_tb) are in green, and MTs in the central region of the spindle body are in cyan (Spindle_midzone). (E) Lifetime of EB1 tracks in the three subspindle regions was fit with a one-phase exponential decay to determine the rate constant K (s −1 ), and the average lifetime (τ) was determined from K −1 . Data represent mean ± 95% confidence interval from the fit (see also Supplemental Figure S2) for 11 spindles for the control with 1045 tracks in aster_lr region, 1063 tracks in aster_tb region, and 4030 tracks in spindle_midzone region and 16 Kif18B RNAi spindles with 3605 tracks in aster_lr region, 2295 tracks in aster_tb region, and 7828 tracks in spindle_midzone region. (F) Histograms of EB1 track velocity in the entire spindle of control and Kif18B knockdowns graphed with the fitting curve of a Gaussian distribution. The mean and SD derived from each fit is indicated. Control RNAi, 11 spindles with 7782 tracks; Kif18B RNAi, 16 spindles with 16414 tracks.
Article Snippet: EB1 comets were automatically detected and frame-to-frame linked using the plusTipTracker MATLAB software package (MathWorks, Natick, MA; Applegate et al. , 2011 ).
Techniques: Knockdown, Transfection, Control, Software, Derivative Assay
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