Figure 1: Spatiotemporal reorganization and coordinated fluctuations of podosome clusters during DC substrate probing. | Nature Communications

Figure 1: Spatiotemporal reorganization and coordinated fluctuations of podosome clusters during DC substrate probing.

From: Actomyosin-dependent dynamic spatial patterns of cytoskeletal components drive mesoscale podosome organization

Figure 1

DCs were transfected with LifeAct-GFP and allowed to adhere on a flat glass coverslip (a) or a coverslip that was scratched with sand paper (see Methods section) (b) and their random slow migration was monitored for several hours at 37 °C by fluorescence microscopy with 3 min frame intervals. Corresponding snapshots at specific times were colour-coded and overlaid. Both on glass and scratches >10 cells from >3 experiments were imaged and one representative movie is shown. DCs were transfected with LifeAct-RFP (c,e) or vinculin-GFP (d,f) and seeded in a glass bottom dish. Imaging was performed at a confocal microscope with 15 s frame intervals. Shown are snapshots of the movies and plots of the RFP/GFP fluctuations over time of three representative neighbouring and distant podosome pairs per condition (grey and black lines). Data are normalized to the maximum intensity. Kymographs (e,f) are shown of the indicated rectangular areas of the LifeAct-RFP and the vinculin-GFP movies. Scale bars 10 μm (a,b,e,f) and 2 μm (c,d).

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