The quantification tool discerns that the effect of cytochalasin B is more pronounced on SFs than on FAs.

<p>(A) Representative images of vinculin in DMSO control cells (top) and Cyt B-treated cells (bottom). (B) Quantification of the number of FAs per cell (a), FAs area (b), and circularity (c) (number of cells: DMSO, n = 33; Cy B, n = 29). (C) (a) The stress fibers quantification tool analyses different parabolic paths (dashed arcs) between two focal adhesions (red ellipses) and selects the parabola that fulfills the intensity requirements (green arc) (Materials and methods). (b) F-actin staining and (c) vinculin staining of a cell after filtering process. (d) Representative example of a z plane containing a stress fiber and the stress fiber in 3D detected by the tool (white line). (e) Image of the stress fibers quantified by the algorithm. (D) Quantification of the number of SFs. (E) Analysis of the number of SFs depending on the number of FAs. (F) Correlation between the number of SFs and FAs. (G) Quantification of the percentage of FAs that have at least one SF (number of cells: DMSO, n = 24; Cyt B, n = 27). Scale bars are 20 µm. ***: p<0.001.