Movie 1.

Flotillin oligomerization induced by a CRY2-CIBN-based optogenetic approach promotes AXL co-clustering. Movie of the boxed area from figure 4B. MCF10A cells that co-express CRY2-mCitrin (green), flotillin 2-CIBN-mCherry (red) and AXL-Halo (blue) labeled with Halo-Tag-Janelia Fluor 646 were imaged by TIRF microscopy. First, flotillin 2-CIBN-mCherry and AXL-Halo were imaged for 8 seconds before 488nm illumination. Then, cells were globally illuminated by 20 pulses (1 pulse/s, 100 ms time length for each) of 488nm-light to stimulate and visualize CRY2. During and after illumination, flotillin 2-CIBN-mCherry and AXL-Halo were continuously imaged. Two examples of flotillin 2/AXL-positive clusters induced by CRY2 oligomerization initiated by illumination are indicated (closed arrow). Co-localization events between flotillin 2-CIBN-mCherry and AXL-Halo are also visible independently of CRY2 oligomerization before illumination and correspond mostly to AXL-containing vesicles positive for flotillin 2 passing through the 200 nm depth of field illuminated by TIRF. Thanks to the high concentration of flotillin 2-CIBN-mCh in these vesicles, CRY2 translocation could be observed. Scale bar: 2 μm.