Fig. 7: The G592R Kv3.3 mutation decreases Hax-1 and increases multivesicular bodies and lysosomes in cerebellar neurons.

Immunoblots and quantification demonstrating a decrease in Hax-1 levels (a n = 4 independent experiments; data are presented as mean ± SEM; two-tailed paired t test), an increase in activated caspase 7 (b n = 5 independent experiments; data are presented as mean ± SEM; two-tailed paired t test), an increase in CD63 levels (c n = 5 independent experiments; data are presented as mean ± SEM; two-tailed paired t test) and an increase in LAMP2 (d n = 4 independent experiments; data are presented as mean ± SEM; two-tailed paired t test) but no change in LC3BII (e n = 3 independent experiments; data are presented as mean ± SEM; two-tailed paired t test) in the cerebellum of G592R Kv3.3 mice compared to those in wild type mice. f Representative electron micrographs of somata of Purkinje cells in wild type and G592R Kv3.3 mice. Red arrows depict lysosomal structures with dark inclusions and blue arrows point to multivesicular bodies, scale bar 2 μm. g Quantification of the numbers of multivesicular bodies (MVB) detected in cross-sections of electron micrographs of somata (left) and of the density of MBVs in the cytoplasm (right) of Purkinje neurons in wild type and G592R Kv3.3 mice. h Representation of proportions of lysosomes either lacking (light) or containing dense inclusions in wild type and mutant mice. i Cytoplasmic density of total lysosomes in wild type and mutant mice. j Quantification of endosomes lacking dense inclusions in wild type and G592R Kv3.3 mice. The data in (f–j) represent measurements from 30 sections each from three wild-type and 3 G592R Kv3.3 mice (data are presented as mean ± SEM; two-tailed unpaired t test). Source data and uncropped Western blots are provided as a Source Data file.