Figure 6: Flux of podosome core and ring components is directionally correlated.

DCs were transfected with LifeAct-RFP and cotransfected with vinculin-GFP (a–c) or talin-GFP (d–f) and seeded in a glass bottom dish. Imaging was performed at a confocal microscope with 15 s frame intervals. Shown are 10 frame moving average images of both channels (a,d). Time series for both channels (100 frames) were subjected to twSTICS, results are shown as vector maps in which the arrows indicate direction of flow and both the size and colour coding are representative of the flow magnitude. Vector maps are plotted onto the immobile filtered version of the image. Shown are the auto-correlation vector maps from the white boxed regions for both channels (b,e). Directional correlation was calculated between vectors measured by twSTICS in two different channels as the cosine of the angle between vectors with the same spatial and temporal lag (c,f). Asterisks indicated statistically significant differences (Mann–Whitney U-test, two-tailed, P<0.001 for c and f). Scale bars, 10 μm.