Figure 2.
Figure 2. Clustering of EB3-GFP growth tracks reveals pVHL’s function as an attenuator of MT shrinkage rate. (A) Clustered EB3-GFP tracks with corresponding head to tail tracks, forward gap closure (gap/pause event), and reverse gap closure (shrinkage event) are shown. A series of tracks and forward gap closures not interrupted by a reverse gap closure is referred to as a growth segment. A circle shows the first detected EB3-GFP signal of a track, and asterisks define the last signal of a track in the corresponding color. Growth track speeds (vm, vo, and vr) are defined as the head to tail distance of the track divided by its lifetime (m, o, and r frames, respectively); gap/pause speed (vn) is defined as the distance between the termination point and the initiation point of the two linked growth tracks divided by the forward gap duration (n frames); accordingly, growth segment speed (vs) is defined by the distance of the entire growth segment divided by the sum of lifetimes of all included growth tracks and forward gap closures (m + n + o frames); shrinkage speed (vp) is defined as the distance between the termination point and the initiation point of the two linked growth tracks divided by the reverse gap duration (p frames). (B) Example of clustered growth tracks in a video acquired from RPE-1 cells. Color codes as in A. (C) Analysis of MT dynamics in n = 12 RPE-1 cells (Fig. 1 A). Distributions of gap/pause speeds of >23,000 forward gap closures (top) and of per cell mean (bottom), distributions of growth segment speeds of >14,000 growth segments (top) and of per cell mean (bottom), and distributions of shrinkage speeds of >10,000 reverse gap closures and per cell mean are shown (***, P < 10−5). (D) Analysis of MT dynamics in n = 10 RCC-4 cells (Fig. 1 B). Distributions of MT gap/ pause speeds of >12,000 forward gap closures (top) and of per cell mean (bottom), distributions of growth segment speeds of >8,000 growth segments (top) and of per cell mean (bottom), and distributions of shrinkage speeds of >5,000 reverse gap closures and per cell mean from 10 cells are shown (*, P < 5 × 10−2; ***, P < 10−5). (C and D) Boxes indicate 25, 50 (median), and 75% quantiles; whiskers extend to 1.5× the interquartile range; red dots indicate outliers beyond this range. Notched boxes indicate uncertainty of the median. Boxes whose notches do not overlap indicate that the medians of the two clusters differ at the 5% significance level. See Materials and methods for statistical test procedures.

Clustering of EB3-GFP growth tracks reveals pVHL’s function as an attenuator of MT shrinkage rate. (A) Clustered EB3-GFP tracks with corresponding head to tail tracks, forward gap closure (gap/pause event), and reverse gap closure (shrinkage event) are shown. A series of tracks and forward gap closures not interrupted by a reverse gap closure is referred to as a growth segment. A circle shows the first detected EB3-GFP signal of a track, and asterisks define the last signal of a track in the corresponding color. Growth track speeds (vm, vo, and vr) are defined as the head to tail distance of the track divided by its lifetime (m, o, and r frames, respectively); gap/pause speed (vn) is defined as the distance between the termination point and the initiation point of the two linked growth tracks divided by the forward gap duration (n frames); accordingly, growth segment speed (vs) is defined by the distance of the entire growth segment divided by the sum of lifetimes of all included growth tracks and forward gap closures (m + n + o frames); shrinkage speed (vp) is defined as the distance between the termination point and the initiation point of the two linked growth tracks divided by the reverse gap duration (p frames). (B) Example of clustered growth tracks in a video acquired from RPE-1 cells. Color codes as in A. (C) Analysis of MT dynamics in n = 12 RPE-1 cells (Fig. 1 A). Distributions of gap/pause speeds of >23,000 forward gap closures (top) and of per cell mean (bottom), distributions of growth segment speeds of >14,000 growth segments (top) and of per cell mean (bottom), and distributions of shrinkage speeds of >10,000 reverse gap closures and per cell mean are shown (***, P < 10−5). (D) Analysis of MT dynamics in n = 10 RCC-4 cells (Fig. 1 B). Distributions of MT gap/ pause speeds of >12,000 forward gap closures (top) and of per cell mean (bottom), distributions of growth segment speeds of >8,000 growth segments (top) and of per cell mean (bottom), and distributions of shrinkage speeds of >5,000 reverse gap closures and per cell mean from 10 cells are shown (*, P < 5 × 10−2; ***, P < 10−5). (C and D) Boxes indicate 25, 50 (median), and 75% quantiles; whiskers extend to 1.5× the interquartile range; red dots indicate outliers beyond this range. Notched boxes indicate uncertainty of the median. Boxes whose notches do not overlap indicate that the medians of the two clusters differ at the 5% significance level. See Materials and methods for statistical test procedures.

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