Supplemental Materials
This article contains the following supporting material:
- Supplemental Materials
- Movie S2 - Movie S2.Actin structures induced by aggregation of VCA diluted with Empty CD16/7 proteins.Movies play in the following order: 100%VCA, 60%VCA, 33%VCA, 14%VCA, 0%VCA. In 60%VCA, 33%VCA and 14%VCA samples, VCA is fluorescently tagged with mCherry (CD16/7-mCherry-VCA) and Empty protein is not tagged with a fluorescent label (CD16/7). In 0%VCA sample, Empty protein is fluorescently tagged with mCherry (CD16/7-mCherry).
- Movie S3B - Movie S3.FRAP experiments and actin structures induced through VCA SynZip binding interface. Movie S3B. Movie of actin aggregates induced by NWASP-VCA undergoing turnover.
- Movie S1 - Movie S1. Actin structures induced by aggregating CD16/7-mCherry-Nck and CD16/7-mCherry-VCA in NIH3T3 cells.
- Movie S6 - Movie S6. Actin pedestals induced by EPEC bacteria in NIH3T3 cells and actin comet tails induced by Vaccinia virus in HeLa cells.
- Movie S4 - Movie S4. Inhibition of formin FH2 domain decreases mobility of Nck-SH3 induced actin assemblies. Movie S4A.Movies and tracks of Nck-SH3 actin structures in cells treated with the 100μM formin inhibitor SMIFH2 (dissolved in DMSO) and in the non-treated control cell (DMSO alone). The treatment/mock-treatment was carried out during acquisition of sixth frame. The movies have 20 frames (every 2.5 min) and total acquisition time is about 49 min. Movie S4B.Movies of Nck-SH3-induced actin structures in cells before and after the treatment with the 150μM formin inhibitor SMIFH2 (dissolved in DMSO) or with DMSO alone (control).Velocity in the SMIFH2 treated cell decreased from 0.026μm/s to 0.01 μm/s. Velocity in the control cell did not change (remained 0.026μm/s after the treatment with DMSO). The movies have 10 frames (every 30 s) and total acquisition time is 4.5 min.
- Movie S3A - Movie S3.FRAP experiments and actin structures induced through VCA SynZip binding interface. Movie S3A. FRAP of CD16/7-mCherry-VCA in clusters and mCherry-VCA in clusters with SynZip binding interface. Scale bars=5μm.
- Movie S5A - Movie S5.Formin FH1 domain overexpression affects dynamics of Nck SH3- but not of the VCA-induced actin structures. Movie S5A. Nck SH3-induced actin structures in the control and in the cells overexpressing FH1 domain of formin mDia1.
- Movie S5B - Movie S5.Formin FH1 domain overexpression affects dynamics of Nck SH3- but not of the VCA-induced actin structures. Movie S5B. VCA induced actin structures in the control and in the cells overexpressing FH1 domain of formin mDia1.