Extended Data Fig. 8: Arf1 and IRSp53 regulate focal adhesion formation and Swip1 and Rab21 regulate cell migration.
From: Cargo-specific recruitment in clathrin- and dynamin-independent endocytosis
(a) Representative immunoblots to validate Swip1 silencing and mScarlet-I-Swip1 or mScarlet-I-∆EF1 overexpression in the experiments shown in Figure 6i. Blot is representative of 3 independent experiments. (b) Representative masks of vinculin-containing focal adhesions in control-, IRSp53- or Arf1-silenced MDA-MB-231 cells and quantification of adhesion number and total area per cell in cells plated on collagen I. Data are presented as mean values ± 95% CI. Statistical significance was assessed with two-sided Mann–Whitney tests, where n is the total number of cells pooled across 3 independent experiments. P values calculated compared to siCTRL condition: ***P = 0.0003, ****P<0.0001. Number of analysed cells over 3 independent experiments: siCTRL, n = 62 cells; Arf1 siRNA #1, n = 60 cells; Arf1 siRNA #2, n = 61 cells; siCTRL, n = 64 cells, IRSp53 siRNA #1, n = 62 cells and IRSp53 siRNA #2, n = 65 cells. Representative immunoblots of 3 independent experiments to validate IRSp53 and Arf1 silencing are shown. Scale bars, 10 µm. (c) Representative phase contrast pictures of the scratch wound after 18 h. Pictures are representative of 3 independent experiments. Quantification of these experiments is shown in Figure 7d. Scale bar, 100 µm. Unprocessed blots and numerical source data are provided in Source data.
