Figure 7. Redeployment of temporal patterning genes generates differentiation trajectories within NB tumors.

(A) Cell trajectory reconstructed from the population of tNBs using semi-supervised pseudotime ordering. Imp⁺ tNBs are enriched at the root of the trajectory while E93+ tNBs are enriched in the branches terminating the trajectories. tNBs are colored according to their State along the trajectory. (B) Dark spline indicates levels of gene expression along the pseudotime. Cells are colored according to their State along the trajectory in (A). p-values for differential expression along the pseudotime are indicated for each gene. (C) Kinetic heatmap depicting the expression of top 200 genes that vary as a function of pseudotime. Genes are grouped in three clusters based on expression kinetics. (D) Gene ontology (GO) enrichment analysis for each cluster identified in (C). (E) Heatmap depicting enrichment of various temporal patterning, metabolic and acetylcholine receptor genes when comparing poxⁿ > pros^RNAi tumors and poxⁿ > pros^RNAi, Syp^RNAi tumors. (F) Gene ontology (GO) enrichment analysis when comparing the transcriptome of poxⁿ > pros^RNAi, Syp^RNAi vs poxⁿ > pros^RNAi tumors.