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Fig. 4 | BMC Biology

Fig. 4

From: The ESCRT-III isoforms CHMP2A and CHMP2B display different effects on membranes upon polymerization

Fig. 4

Mechanical properties of membrane with CHMP protein studied with osmotic shock and HS-AFM indentation. a Confocal images of GUVs submitted to an osmotic pressure difference equal to 150% (osmolarity inside and outside the GUV are respectively 120 mOsm L−1 and 315 mOsm L−1) without (top) or with (bottom) pre-incubation with CHMP2B-ΔC (noted CHMP2B) at a concentration of 1 μM. Scale bar, 10 μm. b Eccentricity index of GUVs (alone or covered with CHMP2B-ΔC polymer) pre-formed in a solution with an osmolarity of 120 mOsm L−1 and transferred to a hypertonic solution with an osmolarity of 315 mOsm L−1 (relative osmotic pressure = 150%). *p-value < 0.05 (Student’s t-test). N = 40. c HS-AFM images of a bare vesicle (left) and a vesicle covered with CHMP2B-ΔC proteins (right). The vesicles with CHMP2B show an increase in surface roughness with respect to the vesicles without CHMP2B (top panels). The deformability of CHMP2B-coated SUVs upon increased applied force is shown at intermediate force increments of 27% (second raw of the panel) and at higher force increment, 81% (third row of panels). d Variation of the relative height of bare vesicles (purple) (N = 31) and vesicles coated with CHMP2B-ΔC (1 μM) (green) (N = 23) or CHMP2B-ΔC (1 μM) + CHMP3 (2 μM) (gray) (N = 30) or MBP-CHMP2A-ΔC (1 μM) + CHMP3 (2 μM) (blue) (N = 69) as a function of force increment. 100% height corresponds to the initial height value, and 0% force increment corresponds to the minimal imaging force. The inverse of the slope of these graphs is directly related to the stiffness. The error bar represents the standard error of the mean (SEM)

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