Fig. 5: AzTax photoswitching allows cell-precise, temporally reversible inhibition of MT polymerisation dynamics. | Nature Communications

Fig. 5: AzTax photoswitching allows cell-precise, temporally reversible inhibition of MT polymerisation dynamics.

From: Photoswitchable paclitaxel-based microtubule stabilisers allow optical control over the microtubule cytoskeleton

Fig. 5

Data related to Supplementary Movies 1 and 2. HeLa cells transfected with EB3-tdTomato treated with 1% DMSO cosolvent with or without 1 µM AzTax3MP. a, b Target cells (violet arrowheads) were selectively illuminated with 405 nm in bursts, and EB3 tracking in defined regions of interest (ROIs) performed. a Target ROIs in frames acquired immediately after 405 nm illumination are outlined in violet, target ROIs in t0 and post-recovery frames (ca. 80 s after 405 nm illumination periods) are outlined in dotted white; nontarget cells are indicated with white arrowheads; scale bars indicate 10 µm. b Quantification of EB3 comet counts (averages over n = 7 cells per treatment group across 3 experiments, each cell’s EB3 comet count time course was normalised to the average of the first ten frames) in target ROI and in non-illuminated cells. Periods of ROI illumination with 405 nm are shaded in violet; times of frames from the representative movies shown in (a) are indicated with dotted lines annotated with the times, data shown as group-average of EB3 comet count time course (normalised to initial count) with s.e.m. For clarity, data for cosolvent-only areas are shown with LOWESS fits. c, d HeLa cells transfected with EB3-tdTomato were treated with either 1 µM AzTax3MP or DMSO cosolvent only, and imaged directly after drug application for 2.4 min with 405 nm illumination (signified by "λ") or without (signified by “d”). Supplementary Movies were analysed for c EB3 comet velocity (n = 12 cells for AzTax3MP; n = 13 cells for cosolvent conditions), and d EB3 comet counts, normalised to cell surface area (n = 13 cells for AzTax3MP lit and both cosolvent conditions; n = 14 cells for AzTax3MP dark condition). EB3 comet velocity could not be meaningfully analysed in AzTax3MP-treated cells under 405 nm illumination due to the strong loss of EB3 comets and is therefore not represented. Data are shown as mean with s.d. Unpaired, two-tailed t test: ***P = 0.001 (95% CI 0.035–0.093), ****P < 0.0001 (95% CI 0.089–0.14), ns denotes not significant, no adjustments for multiple comparisons.

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