JCS02557 Supplementary Material

Files in this Data Supplement:

• Movie 1 - Movie 1. Confocal time-lapse sequence of cell expressing Paxillin-GFP. Time interval between subsequent images: 2 seconds. Focus on cell cortex/substratum interface showing localisation of Paxillin-GFP to filopodia and contact sites that remain stationary once formed.

• Movie 2 - Movie 2. Confocal time-lapse sequence of slug cells expressing Paxillin-GFP. Time interval between subsequent images: 2 seconds. Focus on cortex of cells in the outer cell layer of the slug. Paxillin-GFP localises to many small contact sites, which are generated just behind the leading edge of moving cells. These contact sites remain stationary once formed.

• Movie 3 - Movie 3. 3D reconstruction of prespore cells expressing Paxillin-GFP in an emerging fruiting body. Paxillin-GFP accumulates at cell-cell contact sites. Tilted maximum projection.

• Movie 4 - Movie 4. Confocal time-lapse sequence of cell expressing Paxillin-GFP. Time interval between subsequent images: 5 seconds. Focus on cell cortex/substratum interface. Top left panel: Phase contrast image showing overall cell shape. Top right panel: Reflection interference contrast image in which dark areas correspond to parts of the cell in close proximity to the substratum. The small dynamic almost black structures are part of the contractile vacuole network. Bottom left panel: Paxillin-GFP fluorescence highlighting spot-like contact sites. Bottom right panel: merged fluorescence and interference contrast images. Prior to merging the fluorescence image was inverted to increase the visibility, therefore localised Paxillin-GFP appears purple and the contact area in dark green.

• Movie 5 - Movie 5. Confocal time-lapse sequence of vegetative cell co-expressing Paxillin-GFP and ABD-mRFPmars. Paxillin-GFP (green) localises to tips of filopodia and long-lived spot-like contact sites with the substratum, while the ABD-mRFPmars (red) labels a different set of short-lived contact sites as well as the cortical f-actin and filopodia. Time interval between subsequent images: 3 seconds. Focus on cell cortex/substratum interface.

• Movie 6 - Movie 6. Confocal time-lapse sequence showing a side view of tip/slug formation in an AX2 mound containing 2% mRFPmars labelled AX2 wild-type cells (red) and 2% GFP labelled Paxillin null cells (green). Confocal stacks were acquired every minute. For the movie maximum projections of the fluorescence channels were merged with corresponding bright field images.