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. 2018 Dec 15;7(12):bio038968. doi: 10.1242/bio.038968

Fig. 2.

Fig. 2.

Vero cell spreading morphology. Time is indicated in min. Scale bars: 10 µm. (A) Spreading of untreated cell. (B) Spreading of a cell with stabilized MTs. Cells with stabilized MTs retain the general morphology of normal fibroblasts, although the spreading is mainly anisotropic. (C) Spreading of a cell with depolymerized MTs. Spreading is always anisotropic and substantially decelerated compared to normal fibroblasts. (D) Spreading of a cell with stabilized MTs treated with blebbistatin. Blebbistatin partially restores normal kinetics and morphology of fast spreading in cells with stabilized MTs, in later stages cell exhibits incurved edges, but blebbing disappears compared to cells with stabilized MTs only. (E) Spreading of a cell with depolymerized MTs treated with blebbistatin. Blebbistatin partially restores fast spreading and inhibits blebbing in cells with depolymerized MTs; however, the whole process is relatively slow. On late stages of spreading cell exhibit incurved edges. (F) Spreading of Vero cells in normal conditions (black circles), after stabilization of MTs (black triangles) and after complete depolymerization of MTs (white circles), for each set N=20, data presented as mean±s.e.m. Cells with compromised and stabilized MTs demonstrate lower rates of initial spreading rate and more linear kinetics of spreading process. (G) Spreading of Vero cells in normal conditions (black circles), in the presence of blebbistatin (black triangles) and in the presence of Y-27632 (white circles) for each set N=20, data presented as mean±s.e.m. Myosin II inhibitors do not accelerate early or late spreading. (H) Spreading of Vero cells with completely depolymerized MTs in the presence of myosin II inhibitors (black circles, nocodazole only; white circles blebbistatin and nocodazole; black triangles Y-27632 and nocodazole), for each set N=20, data presented as mean±s.e.m. Myosin II inhibitors partially restore fast spreading kinetics in cells with fully depolymerized MTs. I–Spreading of Vero cells with stabilized MTs in the presence of myosin II inhibitors (black circles–nocodazole and paclitaxel, white circles blebbistatin and nocodazole and paclitaxel, black triangles Y-27632 and nocodazole and paclitaxel), for each set N=20, data presented as mean±s.e.m. Myosin II inhibitors partially restore kinetics of fast spreading (first 10 min) in the cells with stabilized MTs.