JCS020289 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. Podosome dissolution imaged in IRM mode after PGE₂ stimulation. (A) Podosome cores are seen in IRM mode as dark spots. IRM image and the IRM image merged with the vinculin (green) or actin (red) staining. Representative images are depicted. (B) Vinculin is already absent form the podosome rings at the time the bright rings appear, while zyxin is still present. IRM images alone, combined with vinculin (green) or zyxin (green) and/or actin (red) and vinculin (green) or zyxin (green) combined with the actin (red) staining are shown. Representative images are depicted. (C) Myosin IIA localizes to the areas of the bright rings. IRM images alone or combined with myosin IIA (green) and/or actin (red) staining or myosin IIA (green) combined with actin (red) staining. Representative images are depicted. The white arrow indicates a region with bright rings and prominent myosin IIA staining.

• Movie 1 -

  Movie 1. PGE₂-induced podosome loss and FA formation and contraction of the DC. A combination of DIC images (left) and IRM images (right) show an iDC displaying podosomes. Shortly after stimulation with PGE₂ the podosome rings (transiently) light up just prior to podosome dissolution, while FAs are formed at the border of the cell. Stills from this video can be seen in Fig. 2B. Images were collected every 20 seconds and the display rate is 9 frames/second.

• Movie 2 -

  Movie 2. PGE₂-induced podosome loss and FA formation and contraction of the DC. A combination of DIC images (left) and IRM images (right) show an iDC displaying podosomes. Shortly after stimulation with PGE₂ the podosome rings (transiently) light up just prior to podosome dissolution, while FAs are formed at the border of the cell. Images were collected every 20 seconds and the display rate is 9 frames/second.

• Movie 3 -

  Movie 3. PGE₂-induced podosome loss is blocked by myosin II inhibition in DCs. A combination of DIC images (left) and IRM images (right) show an iDC displaying podosomes in the presence of blebbistatin. Upon stimulation with PGE₂ the podosomes stay intact. Images were collected every 20 seconds and the display rate is 9 frames/second.