Fig. 1: Giant Organelles Vesicles’ recovery and characterization.

a Organelles reshaping into Giant Organelle Vesicles (GOVs) by a hypotonic medium. Confocal microscopy image of a swollen COS-7 cell expressing RFP-KDEL and Mfn2-YFP. Confocal microscopy snapshot showing the release of Giant ER Vesicles (GERVs) after plasma membrane rupture with a micropipette. Cells expressing GPI_2x-mCherry (Plasma Membrane) and RFP-KDEL(ER). Scale bar: 5 µm. Schematic representation of a collected GOV. b Diameter distribution of Giant Organelle Vesicles (GOV). Only vesicles larger than 0.75 µm were measured. For each organelle type, quantifications were done from two independent experiments with hundreds of vesicles quantified from tens of cells. c Confocal images of collected Giant Organelle Vesicles (GOVs) labeled with fluorescence markers representing different organelles. Scale bars: 5 µm. d Plot illustrating the initial membrane tension of Giant Organelle Vesicles (GOVs). For organelles from left to right, the number of independent experiments is N = 6; N = 4; N = 8; N = 4; N = 4; N = 4. Each experimental data point corresponds to a GOV from a different cell and the mean of each replicate is shown. Nested One-Way ANOVA - Multiple comparisons: Fisher LSD test. Individual P values are shown. e The lysis tension of Giant Organelle Vesicles (GOVs). For organelles from left to right, the number of independent experiments is N = 4; N = 4; N = 7; N = 4; N = 4; N = 4. Each experimental data point corresponds to a GOV from a different cell and the mean for each replicate is shown. Nested One-Way ANOVA - Multiple comparisons: Fisher LSD test. Individual P values are shown. f The apparent area expansion modulus of Giant Organelle Vesicles (GOVs). For organelles from left to right, the number of independent experiments is N = 4; N = 6; N = 3; N = 3. Each experimental data point corresponds to a GOV from a different cell and the mean for each replicate is shown. Nested One-Way ANOVA - Multiple comparisons: Fisher LSD test. Individual P values are shown. g Bending rigidity of Giant Organelle Vesicles (GOVs). For organelles from left to right, the number of independent experiments is N = 2; N = 3; N = 3; N = 3, N = 3. Each experimental data point corresponds to a GOV from a different cell and the mean for each replicate is shown. Nested One-Way ANOVA - Multiple comparison: Fisher LSD test. Individual P values are shown.