Fig. 5: Differentiation of SCA3 and isogenic control SCA3-NSCs into cerebral cortical neurons.

a–c Immunofluorescence was used to detect the expression of surface markers and synaptic proteins in each group. Further differentiation resulted in neurons expressing TUJ1, MAP2, GABA, and GFAP. e and f The percentage of corresponding neuron markers were counted. Similar results were acquired with each cell line in our study (P > 0.05). Scale bar (TUJ1): 50 µm. Scale bar (MAP2, GABA, and GFAP): 25 µm. d, g Representative images of neuronal cells stained with SYP1 and PSD95 on days 35–50, there was no difference in the expression of SYP1/PSD95 in each group (P > 0.05). Scale bars 25 µm. h IC2 positive polyQ aggregates were detected in SCA3 neurons compared with other groups on day 50 (aggregates indicated in enlarged images with the arrow). Scale bar: 100 µm. b, d, f, h All data were presented as mean ± SD. The data as determined by one-way ANOVA and Bonferroni post hoc test. n = (6, 6, 6, 6, 6, 6) images for TUJ1, MAP2, GABA, GFAP, SYP1, and PSD95 neuronal markers expression. Ctrl-NCs: healthy control neurons. SCA3-NCs: SCA3 patient-derived neurons. SCA3-C3-NCs and SCA3-C12-NCs: corrected SCA3 neurons.