Fig. 4
Correlated Live Microscopy-Electron Tomography of Ultrastructural Changes in Networks with Increasing Filament Density (A) A migrating keratocyte expressing lifeact:GFP was aspirated at the rear with a micropipette, fixed within ≈3 s, and prepared for electron microscopy. The cell shifted out of focus because of the manipulating procedure. A transient increase in actin density can be seen in the low-magnification electron micrograph on the right. (B) 5.5-nm tomogram slice of the region marked by a red box in (A). The rear of the cell is toward the left side of the picture, and the cell front is seen on the right. Regions of steady-state density, increased density, and decreased density are marked with black, blue, and red throughout the whole figure. (C) Filament tracks of the lamellipodium shown in (B), with actin filaments shown in green, barbed ends in red, and pointed ends in blue. (D) Filament numbers and densities of barbed and pointed ends in 106-nm-wide spatial bins throughout the lamellipodium shown in (B) and (C). (E) Histogram showing filament angles to the cell edge in 212-nm distance bins. See also Figures S4C and S4D and Movie S3. |
Reprinted from Cell, 171, Mueller, J., Szep, G., Nemethova, M., de Vries, I., Lieber, A.D., Winkler, C., Kruse, K., Small, J.V., Schmeiser, C., Keren, K., Hauschild, R., Sixt, M., Load Adaptation of Lamellipodial Actin Networks, 188-200.e16, Copyright (2017) with permission from Elsevier. Full text @ Cell