Figure 7

CORO2B increases CFL1 recruitment to focal adhesion sites and actin fibers. (a) Schematic illustrating the generation of a SILAC based quantitative CORO2B dependent focal adhesome by expression of CORO2B or GFP as control in human podocytes with subsequent focal adhesion isolation and mass spectrometry analysis. (See Methods section and Supplementary Dataset S1.) (b) Mapping of enriched and reduced proteins reveals impaired recruitment of the integrin receptors dimer ITGB1/ITGAV to CORO2B dependent focal adhesions. In contrast CORO2B and CFL1 recruitment to focal adhesion sites was increased. (c) CFL1 co-precipitates with CORO2B. V5-tagged CORO2B was immunoprecipitated by anti-V5 coated Sepharose beads. (d–f) Localization studies of fluorophore tagged CFL1 and CORO2B or PAXILLIN as control reveals CORO2B dependent recruitment of CFL1 to focal adhesions and fibrillary actin bundles. F-ACTIN was visualized by Phalloidin staining. Dotted lines indicate regions for line scan measurements of fluorescence intensities.