Figure 1.

F-actin flow characterized using TIRF-SIM and STICS analysis. (a) Images show characterization of F-actin flow in T cells; shown are representative TIRF-SIM images (a, e, i, and m) of cells 5–10 min after contact with a stimulatory coverslip and after different treatments (labeled bottom left of images). ROIs within the blue boxes are to the right (b, f, j, and n), showing the output vector maps, from STICS analysis, representing ≈1 μm². Scale bars, 5 μm. (b) Graphs show STICS outputs of normalized F-actin flow; any changes after drug treatment in either subregion velocity (c, g, k, and o) or directionality (d, h, l, and p) are plotted. Vector angles are calculated against a seed point (cell center) as shown in (a) as a red dot, where a vector oriented toward a seed point gave an angle of 0° (solid arrow), perpendicular to 90° (dotted arrow), and away from 180° (dashed arrow). (Controls: 1.63 ± 0.46 μm, n = 24. Cytochalasin-D (C-D): 0.99 ± 0.27 μm, n = 10. Jasplakinolide (Jasp): 2.07 ± 1.06 μm, n = 27. 7-ketocholesterol (7KC): 1.61 ± 0.49 μm, n = 24.) To see this figure in color, go online.