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. 2021 Apr 19;32(9):869–879. doi: 10.1091/mbc.E20-11-0723

FIGURE 2:

FIGURE 2:

Intensity difference–based measurement of polymerization and depolymerization rates in the interior and the periphery of microtubule asters. (A) TIRF microscopy movies of fluorescent tubulin were subjected to intensity difference analysis, which revealed a spatial map of polymerization and depolymerization for the interior (<100 µm away from the MTOC) and peripheral (>200 µm from the MTOC with few microtubules) region of microtubule asters. These images correspond to Supplemental Movies 2 and 3. (B) The distributions of polymerization and depolymerization rates measured from N = 6 interior movies and N = 6 peripheral movies. The polymerization rate (median 32.0 vs. 32.0 µm/min) showed no statistical difference in the interior vs. periphery (p-value = 0.819, t test). The depolymerization rate (median 37.6 vs. 26.7 µm/min) was higher in the interior (p-value = 4.44e-16, two sample Kolmogorov-Smirnov tests) with estimated difference of mean and median depolymerization rates of 7.1 µm/min (95% CI = [5.9, 8.3]) and 11.0 µm/min (95% CI = [8.4, 12.4]). Vertical lines indicate the mean for each distribution. See Supplemental Figure 1 for analysis of individual movies. Scale bar, 20 µm.