ESCRT structures surround actin-independent integrin clusters. (A) Confocal micrographs of monocyte-derived dendritic cells (moDCs) incubated for 40 min and immunolabeled for IST1 (magenta in merge), integrin αM or integrin β2 (cyan), and phalloidin (yellow). Blue: DAPI. Bar graphs show cells with integrin-wrapping IST1 structures as a percentage of all cells forming IST1 structures, at the indicated time points, for integrin β2 and integrin αM. Line graphs show fluorescence intensity profiles as indicated by the white line. (n ≥ 3 donors; two-sided paired t tests; *: P < 0.05; **: P < 0.01; NS: not significant). Scale bars: 10 µm. (B) Immunofluorescence labeling of a temporal artery biopsy (same donor 1 as in Fig. 5 A) for DAPI (blue), IST1 (magenta), integrin β2 (yellow), and macrophage marker CD68 or fibroblast marker vimentin (cyan). (C) Confocal micrographs of moDCs seeded in the presence or absence of 5 mM EDTA, and immunostained for IST1 (magenta), integrin β2 (green), and phalloidin (yellow). Arrows: IST1-positive structures. Graph: average number of IST1-positive structured per cell (n = 3 donors). For the statistical analysis of A and C, data distribution was assumed to be normal, but this was not formally tested. Scale bars: 10 µm.