Figure 5: The diameter of DNM2-V96 structures decreases with temperature and inhibition of Dynamin GTPase activity.
HEK393T cells were transiently transfected and incubated with or without dynasore or at different temperatures. Dynasomes were stained using secondary immunofluorescence (white) and imaged using confocal microscopy. The diameters for a large dataset of these structures were estimated using image analysis. Nuclei were stained with DAPI (Blue). A) At 37°C Dynasome-S morphologies are predominant. B) After incubation with 80 μM dynasore, the size of vesicular structures collapses. C) As temperature increases, the mean diameter of dynasome morphologies decreased significantly (****p<0.0001, ANOVA, Tukey post hoc). D) Having validated that this image analysis reflects the trend observed with temperature, this approach was used to compare the diameter of structures with and without dynasore. Dynasore treatment significantly decreased the diameter (p < 0.0001, two-tailed unpaired t-test).