Fig. 1
EDPs cause fast membrane bleb formation in HT-1080 cells. a Scanning electron microscopy of control (Ctl) and EDP-treated HT-1080 cells after 40 min of incubation. Scale bar: 10 µm. b EDP dose-dependent induction of blebbing was evaluated by counting 10 fields per well under a phase contrast optical microscope after a 40 min incubation. Results were expressed as mean ± S.E.M. Data from one experiment, representative of three independent experiments, are shown. c HT-1080 cells were treated with 50 µg/mL EDPs from 0 to 120 min. The quantification of the blebbing cell number was evaluated by counting 10 fields per well under a phase contrast optical microscope. Results were expressed as mean ± S.E.M. Data from one experiment, representative of three independent experiments, are shown. d EDPs stimulate cell membrane blebbing in 3D collagen matrix. GFP-Hsp90 transfected HT-1080 cells were seeded in a 3D collagen matrix in the presence of EDPs. 3D confocal imaging was realised after 24 h of incubation to study the HT-1080 cell morphology. In the presence of EDPs, HT-1080 cells adopted an amoeboid morphology with many membrane blebs and shed extracellular vesicles (white arrows). Scale bar: 10 µm. e Real-time imaging and analysis of bleb formation in GFP-Hsp90-transfected cell. Live-cell imaging by microscope coupled with confocal Yokogawa spinning disk of control and EDP-treated HT-1080 cells. f Analysis of bleb expansion and retraction on cell sections (red line Y-axis) on the 3D real-time sequences. Snapshots of the time series at 12 s intervals from which the analysis were generated (Z-axis/Y-axis). Data from one experiment, representative of 10 independent experiments are shown. g Graph of a bleb extension and retraction following the cell section analysis. Results are means of a representative experiment (n = 10) repeated on five independent experiments. h Snapshots of time-lapse phase-contrast microscopy movies of HT-1080 blebbing cell in presence of EDPs. Scale bar: 10 µm. *p < 0.05, **p < 0.01, ***p < 0.001, significantly different from control
