Figure S3. Non-endocytic clathrin... | Rockefeller University Press

Figure S3.

$Non-endocytic clathrin structures. Results of LocMoFit analysis of clathrin structures in SK-MEL-2 cells (N = 13 cells, n = 1,798 sites). (A) A strong correlation between curvature and θ can be observed for most structures (n = 1,645 sites, black). A disconnected point cloud (n = 153 sites, 8.5%, red) indicates the presence of endocytosis-unrelated clathrin structures. (B–D) The same distinct population of data points can be observed for the (B) surface area, (C) edge length, and (D) projected area. (B) Example structures from the disconnected population of sites in top view (xy) and 50 nm-thick z-slices (xz) and their respective fitted θ values. Scale bar, 100 nm. (C–F) Analysis of clathrin coats not following general trajectory of curvature generation. (C) 3T3 cell transiently overexpressing AP2-GFP. Left: Single-molecule localization microscopy image of immunolabeled clathrin. Middle: Diffraction-limited image of the AP2-GFP signal. Right: Overlay of the two targets (scale bar, 10 µm). (D) Enlarged image of the section indicated in C (scale bar, 1 µm). (E) Example sites indicated in B (scale bar, 100 nm). 1: Example for a structure annotated as “GFP positive.” 2: Example for an “inconclusive” GFP signal. 3 and 4: Example of “GFP negative” structures.” (F) Analysis results when estimating θ and curvature from clathrin structures and annotating them depending on their AP2 signal (N = 3 cells and n = 277 sites). No AP2-GFP positive structures are found in the disconnected population of sites, suggesting that they are most likely not generated via clathrin-mediated endocytosis.$

Non-endocytic clathrin structures. Results of LocMoFit analysis of clathrin structures in SK-MEL-2 cells (N = 13 cells, n = 1,798 sites). (A) A strong correlation between curvature and θ can be observed for most structures (n = 1,645 sites, black). A disconnected point cloud (n = 153 sites, 8.5%, red) indicates the presence of endocytosis-unrelated clathrin structures. (B–D) The same distinct population of data points can be observed for the (B) surface area, (C) edge length, and (D) projected area. (B) Example structures from the disconnected population of sites in top view (xy) and 50 nm-thick z-slices (xz) and their respective fitted θ values. Scale bar, 100 nm. (C–F) Analysis of clathrin coats not following general trajectory of curvature generation. (C) 3T3 cell transiently overexpressing AP2-GFP. Left: Single-molecule localization microscopy image of immunolabeled clathrin. Middle: Diffraction-limited image of the AP2-GFP signal. Right: Overlay of the two targets (scale bar, 10 µm). (D) Enlarged image of the section indicated in C (scale bar, 1 µm). (E) Example sites indicated in B (scale bar, 100 nm). 1: Example for a structure annotated as “GFP positive.” 2: Example for an “inconclusive” GFP signal. 3 and 4: Example of “GFP negative” structures.” (F) Analysis results when estimating θ and curvature from clathrin structures and annotating them depending on their AP2 signal (N = 3 cells and n = 277 sites). No AP2-GFP positive structures are found in the disconnected population of sites, suggesting that they are most likely not generated via clathrin-mediated endocytosis.