Figure 6.

Age-dependent neurodegeneration in TECPR2^−/− mice. (A) Age-dependent progressive formation of spheroids in the tecpr2^−/− mouse brainstem. Tecpr2^+/+ and tecpr2^−/− mice (n = 3 for each experiment) aged 3, 6 and 9 months were perfused with 4% PFA and their brains were then removed and embedded in paraffin. Coronal brain sections obtained from similar areas of the brainstem were stained with H&E and analyzed by widefield microscopy. Number of spheroids was normalized to the size of the measured area (right panel). Scale bar of upper panel: 500 µm, scale bar of zoomed images: 50 µm. *p < 0.05, **p < 0.01 determined by one-way ANOVA with Dunnett’s Multiple Comparison Test as posttest. (B) Age-dependent progressive accumulation of LC3 in large autophagosomal structures in the brain affected regions of tecpr2^−/− mice. Coronal brain sections from brainstem regions, corresponding to spheroid areas in mice aged 3, 6, and 9 months, were immunostained for LC3 (green) and analyzed by confocal microscopy. Scale bar: 20 µm. The intensity of LC3B was calculated by ImageJ and presented with the SEM * p < 0.05, **p < 0.01 determined by t-test (right panel)