Fig. 4

Identification of neural progenitors in the VNC. A Dot plot showing expression of cluster 10 marker genes within cluster 10 and across all other clusters. B Cell-cycle phase analysis showing the percentage of cells in G1, G2/M, and S phase, assigned according to marker gene expression. *P < 0.05, Pearson’s Chi-square test with a post-hoc Bonferroni Adjustment for multiple comparisons. (C-F) Identification of neuroblasts, intermediate progenitors, and differentiating neurons. C Seurat UMAP plot depicting 5 cluster 10 sub-clusters. D Dot plot depicting marker gene expression for cluster 10 sub-clusters. Neurotransmitter marker genes were highlighted by bold font. E Cell-cycle phase analysis of cluster 10 sub-clusters. Sub-cluster 4 had a significantly larger portion of cells in G2M phase than other clusters and had no cells expressing G1 markers. *P < 0.05, Pearson’s Chi-square test with a post-hoc Bonferroni Adjustment for multiple comparisons. F Pseudo-time trajectory analysis of neural progenitor cells from cluster 10, anchored at sub-cluster 4. Cells are colored according to a lookup table that depicts their pseudo-time score along the trajectory, with the darkest color indicating the most-immature neurons. G Identification of mitotic populations in the VNC. Top, schematic depicting the workflow for EdU incorporation experiments. Bottom, representative maximum intensity projection images depict EdU immunoreactivity in thoracic and abdominal ganglia. H Plots depict the number (left) and fraction of cells in the indicated segments (T1, A1, A4, A7) which incorporated EdU. Boxes represent mean values, bars represent standard deviation, and points represent measurements from individual segments. *P < 0.05, Kruskal-Wallis test with a post-hoc Dunn’s test. Genotype: wild type in (G, H); all other panels: brp-T2A-QF2w / +; QUAS-mcd8GFP / +