Extended Data Fig. 3: SIM images of Swip1 with CG components and Rab proteins.
From: Cargo-specific recruitment in clathrin- and dynamin-independent endocytosis
(a) Confocal micrographs of MDA-MB-231 cells expressing mScarlet-I-Swip1 and GFP–Rab21, and immunostained for β1-integrin (12G10 antibody). Arrows show areas of co-localization in endosomal structures. Representative pictures of n = 3 independent experiments. Scale bar, 10 µm. (b) Full pictures from Fig. 2d, e and 3d. x-y projections of MDA-MB-231 cells expressing mScarlet-I-Swip1 and either GFP–Rab21, Arf1–GFP or GFP–IRSp53 were imaged using structured illumination microscopy (SIM). Blue and yellow squares highlight the regions of interest (ROI) shown as x-z projections in Fig. 2d, e and Figure 3d. Representative pictures of n = 3 independent experiments. Scale bars, 5 µm (2d), 8 µm (2e) and 8 µm (3d); insets, 1 µm. (c) SIM x-z projections of MDA-MB-231 cells expressing mScarlet-I-Swip1 and immunostained for endogenous Rab proteins and quantification of Rab protein co-localization with mScarlet-I-Swip1. Each dot represents the co-localization ratio in one cell. 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 from 3 independent experiments. P values calculated compared to Rab21 condition. **** P < 0.0001. Number of cells analysed over 3 independent experiments: n = 39 cells for Rab21, n = 35 for Rab5, n = 31 for Rab7, n = 36 for Rab11. Scale bars, 0.5 µm. Numerical source data are provided in Source data.
