Fig. 5: Confocal microscopy analysis and classification of the Httex1 inclusions formed in neurons revealed their morphological heterogeneity.

a Httex1 expression was detected by ICC staining combined with confocal imaging in primary cortical neurons, 3 (D3), 7 (D7), and 14 (D14) days after lentiviral transduction. Httex1 mutants were detected with the MAB5492 antibody. The nucleus was counterstained with DAPI (blue), and MAP2 was used to visualize the neurons (red). Scale bars = 20 μm. b Image-based quantification of the number of neurons containing Httex1 inclusions over time. The graphs represent the mean ± SD of 3 independent experiments. c Primary neurons were classified via the detection of Httex1 as diffuse or by the morphology of the detected Httex1 aggregates: (1) Small nuclear puncta; (2) at least one large nuclear inclusion, and (3) cytoplasmic inclusion. In addition, a subclass was created for neurons containing a large nuclear inclusion associated with nuclear condensation. Scale bar = 20 μm. d Image-based quantification and classification of the different morphologies of Httex1 inclusions based on the panel c. A minimum of n = 145 cells per condition were examined over 3 independent experiments. Data are presented as mean values + /− SD. Statistical analysis: One-way ANOVA followed by a Tukey [HSD] post hoc test was performed. p-values < 0.0001 for top and bottom panels and *P < 0.05, **P < 0.005, ***P < 0.001 for multiple comparisons. e Image-based quantification of neurons containing a large nuclear inclusion with a nuclear condensation. A minimum of n = 145 cells per condition were examined over 3 independent experiments. Data are presented as mean values + /− SD. Statistical analysis: Two-way ANOVA revealed no significant interaction (p-value = 0.0598) but a significant row factor samples (p-value = 0.0076) and column factor time (p-value = 0.0002) with *P < 0.05, **P < 0.005, ***P < 0.001 for multiple comparisons.