Fig. 6: Induced neurons carrying different SETBP1 variant groups showed distinct differentiation capacity and morphology, and transcriptomic profiles. | Nature Communications

Fig. 6: Induced neurons carrying different SETBP1 variant groups showed distinct differentiation capacity and morphology, and transcriptomic profiles.

From: SETBP1 variants outside the degron disrupt DNA-binding, transcription and neuronal differentiation capacity to cause a heterogeneous neurodevelopmental disorder

Fig. 6

a Schematic representation of the SETBP1 protein (UniProt: Q9Y6X0) indicating the locations of variants included. b Schematic representation of the generation of induced neurons from fibroblasts. Asterisks indicate the time of harvest. CA: compound set A; CB: compound set B. c Confocal microscopy images of immunostained neuronal markers Tuj1 (green), MAP2 (orange), and DCX (magenta) in control and patient-derived induced neurons. Nuclei were stained with Hoechst 33342 (white). Merged images are shown. Results are representative of three independent experiments. Scale bar = 100 μm. d Quantification of the proportion of Tuj1-positive cells (%Tuj1/Hoechst) at days 10 and 12. Bars represent the mean ± SEM of three independent experiments (vs controls; one-way ANOVA and a post-hoc Dunnett’s test). Details of statistical tests and p-values are provided in Source Data 9. e Zoomed in confocal microscopy images of immunostained neuronal markers Tuj1 (green) in control and patient-derived induced neurons. Nuclei were stained with Hoechst 33342 (white). Results are representative of three independent experiments. Scale bar = 50 μm. f Quantification of neuronal morphology using Sholl analysis. Violin plots of three independent experiments (vs controls; one-way ANOVA and a post-hoc Dunnett’s test). Brown crossbars indicate the median. At least 40 neurons per cell line from three independent differentiation experiments were measured. Details of violin plot statistics, statistical tests and p-values are provided in Source Data 9. g Sholl profile of neuronal morphology. Data are from three independent experiments. Individual data points represent the number of dendritic intersections per radial distance from the soma. Solid lines show the predicted mean per condition, fitted using cubic polynomial regression (3rd-degree) via linear modelling; shaded bands represent 95% confidence intervals around the model fit. Statistical analysis was performed using one-way ANOVA and a post-hoc Tukey’s test. At least 40 neurons were analysed per cell line. Details of statistical tests and adjusted p-values are provided in Source Data 9. Source data are provided as a Source Data file.

Back to article page