Fig. 3

TMEM106B overexpression induces lysosomal dysfunction. A Representative images of hTMEM106B(+) and wild-type neurons stained for LAMP1 and TMEM106B, acquired with the same acquisition parameters, and displayed with identical brightness and contrast settings. Asterisks indicate example neurons that were included in the intensity measurement by discrimination from glial and/or dead cells based on nuclear features (see materials and methods for details). Scale bars (50 µm) and B Intensity measurements in wild-type and hTMEM106B(+) neurons show higher TMEM106B and LAMP1 intensity in TMEM106B overexpression neurons. Datapoints represent the average intensity per replicate (coverslip). Axonal lysosomal transport was evaluated by live cell imaging using Lysotracker. There was no significant difference in lysosomal mobility or speed between wild-type and hTMEM106B(+) neurons, quantified in C-D. Datapoints represent the average mobility or speed of tracks in one image field. Data represented as mean ± SEM. T-test *, P < 0,05; **, P < 0,01; ***, P < 0,001; ****, P < 0,0001