Analysis of endocytosis spatial organization.

<p>(A) Clathrin-independent IL-2R putative endocytic sites. Top: IL-2R is labeled with fluorescent antibodies and imaged using total internal reflexion fluorescence (TIRF) microscopy. Bottom: We delimited individual cells by drawing polygonal (green) Regions of Interest (ROIs) in the software Icy <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080914#pone.0080914-deChaumont1" target="_blank">[13]</a> (<a href="http://icy.bioimageanalysis.org" target="_blank">http://icy.bioimageanalysis.org</a>). Positions of putative endocytic sites (objects) inside each cell are then extracted with a multi-scale wavelet analysis <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080914#pone.0080914-OlivoMarin1" target="_blank">[28]</a>. (B) The spatial organization of IL-2R putative endocytic spots is quantified with (solid black line). CF expansion of and is represented with black dotted lines. In the bottom-right corner, the mean statistic (Eq. 8) is plotted against ( cells, objects)). (C) Clathrin putative endocytic sites. Top: Clathrin light chain is fused with green fluorescent protein (GFP) and imaged using TIRF microscopy. Bottom: Positions of putative endocytic sites are extracted with a multi-scale wavelet analysis <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0080914#pone.0080914-OlivoMarin1" target="_blank">[28]</a>. (D) The spatial organization of clathrin putative endocytic spots is quantified with (solid black line). CF expansion of and is represented with black dotted lines. In the bottom-right corner, the mean statistic (Eq. 8) is plotted against ( cells, objects). Scale bar = 5 microns.