Fig. 9: The Kv3.3 G592R mutation promotes release of exosomes into the extracellular space.

a Representative transmission electron microscopy photomicrographs of low density fraction (LDF) and intermediate-density fraction (IDF) extracellular vesicles (EVs) isolated from the cerebellum of a wild type mouse. Scale bar: 500 nm. Images are representative of ten images taken of each of the sets analyzed. b Representative Western blot analyses of LDF EVs (L) and IDF EVs (I) compared to the cerebellar homogenate (BH). c Nanotrack analysis (NTA) of the hydrodynamic diameter of the EVs found in LDFs and IDFs. The curve is normalized to the mode of each distribution (five independent isolations. d Quantification of particles in LDFs and IDFs by NTA that are found within the size bins shown in the graph. Data are plotted as percentage of total number of EVs. Five independent isolations. e Representative Western blot analyses of EVs isolated from cerebella of wild type and G592R Kv3.3 mutant mice. LDFs (L) and IDFs (I) were isolated from cerebella of two mice of the same genotype and these were combined and tested for the markers shown. Two independent experiments are shown in the blot. f–h quantification of data in e. The densitometric quantification was performed on three independent experiments, representative of six mice per genotype. BCA protein assay (i) and total particle count as estimated by NTA (j) of LDFs and IDFs in wild type and G592R Kv3.3 mice. Values were normalized to the cerebellum wet weight (five independent isolations for wild type and three independent isolations for mutant mice, corresponding to ten and six mice, respectively. All data are shown as a mean ± SEM. Statistical test used in all figures: Two-Way ANOVA with Sidak’s multiple comparisons test). Source data and uncropped Western blots are provided as a Source Data file.