Figure S3.
Characterization of the PIV analysis, the effects of latrunculin B treatment on the organization of filamentous actin, and extended Fourier analysis of unperturbed microtubules, related toFigs. 2 and 3. (a–d) The resolution of the PIV analysis was evaluated by performing the same analysis for control cells which were not subjected to external forces. The analysis was performed for standard confocal (a and c) and super-resolution confocal (b and d). (a and b) Left: Two successive snapshots of live CHO cells expressing EGFP-tubulin were acquired at 1 sec interval and analyzed by PIV. The result shows that the detected displacements were significantly small compared to the cells that applied creep forces for the same 1 sec (shown in c and d). Right: Probability of the displacements detected in control. The probability distribution was well fitted with a single Gaussian function with the variance σ= 0.019 ± 0.002 μm (standard confocal) and σ= 0.015 ± 0.001 μm (super-resolution confocal). (c and d) Representative single-cell data of the deformation analysis. The local displacement of microtubules of a single cell in a creep experiment is shown with the value of σ. The plot indicates that most of the detected displacements in the creep experiment are above the noise level. Scales are 10 μm for the microscopy image (white bar) and 0.2 μm for the displacement vector (green arrow). (e) Orientational analysis of the deformation of microtubules. The angle of microtubule displacement is plotted as a function of the predicted angle from the model. The distance from the load position and the amplitude of the displacement are color coded. (f and g) The filamentous actin of control (f) and latrunculin B-treated (g) cells were visualized by Lifeact-StayGold. The actin meshwork observed in control cells was not observed in latrunculin B-treated cells. The imaging condition and image contrast are the same for the two conditions. Scale bars are 10 μm (left) and 2 μm (right enlarged and enhanced). (h and i) Fourier analysis of unperturbed microtubule shape using standard (h) or super-resolution (i) confocal microscope. Left: Representative snapshots of microtubules visualized using Tubulin-EGFP. Scale bar is 10 μm. Right: The ensemble-averaged square amplitude of the Fourier modes of microtubule shape plotted as a function of the wavenumber. Broken line indicates slope −2.

Characterization of the PIV analysis, the effects of latrunculin B treatment on the organization of filamentous actin, and extended Fourier analysis of unperturbed microtubules, related to Figs. 2 and 3. (a–d) The resolution of the PIV analysis was evaluated by performing the same analysis for control cells which were not subjected to external forces. The analysis was performed for standard confocal (a and c) and super-resolution confocal (b and d). (a and b) Left: Two successive snapshots of live CHO cells expressing EGFP-tubulin were acquired at 1 sec interval and analyzed by PIV. The result shows that the detected displacements were significantly small compared to the cells that applied creep forces for the same 1 sec (shown in c and d). Right: Probability of the displacements detected in control. The probability distribution was well fitted with a single Gaussian function with the variance σ= 0.019 ± 0.002 μm (standard confocal) and σ= 0.015 ± 0.001 μm (super-resolution confocal). (c and d) Representative single-cell data of the deformation analysis. The local displacement of microtubules of a single cell in a creep experiment is shown with the value of σ. The plot indicates that most of the detected displacements in the creep experiment are above the noise level. Scales are 10 μm for the microscopy image (white bar) and 0.2 μm for the displacement vector (green arrow). (e) Orientational analysis of the deformation of microtubules. The angle of microtubule displacement is plotted as a function of the predicted angle from the model. The distance from the load position and the amplitude of the displacement are color coded. (f and g) The filamentous actin of control (f) and latrunculin B-treated (g) cells were visualized by Lifeact-StayGold. The actin meshwork observed in control cells was not observed in latrunculin B-treated cells. The imaging condition and image contrast are the same for the two conditions. Scale bars are 10 μm (left) and 2 μm (right enlarged and enhanced). (h and i) Fourier analysis of unperturbed microtubule shape using standard (h) or super-resolution (i) confocal microscope. Left: Representative snapshots of microtubules visualized using Tubulin-EGFP. Scale bar is 10 μm. Right: The ensemble-averaged square amplitude of the Fourier modes of microtubule shape plotted as a function of the wavenumber. Broken line indicates slope −2.

This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal