Figure 1: Myosin IIA-mediated contractility is not essential for podosome core integrity.

(a) Myosin IIA is associated with FAs and podosomes. Immature DCs were seeded on glass coverslips, fixed and stained with Texas Red-conjugated phalloidin and a fluorescent anti-myosin IIA mAb to visualize actin and myosin IIA, respectively. Squares represent enlarged views of typical FAs and podosome (pod) areas. Scale bar, 10 μm. (b) DCs were transfected with either VinTS or VinTL and fluorescence lifetime was measured before and after 30 min of treatment with 20 μM blebb. Shown is the frequency distribution of the fluorescence lifetime of at least 800 individual podosome rings in 20 different cells and the Gaussian fit of the data. (c) Immature DCs were seeded on glass coverslips and left untreated or stimulated with 20 μM blebb for 5, 10, 20 and 30 min. Cells were subsequently fixed and stained with Texas Red-conjugated phalloidin to quantify the number of podosome-bearing cells and the FA markers vinculin, talin, zyxin and paxillin to quantify the number of FA-bearing cells. Mean±s.d. of the percentage of cells displaying podosomes or FAs for at least 100 cells per condition is shown. Statistical analyses were performed by two-tailed Student’s t-test. **P<0.01, ***P<0.001.