Figure 4: SCENT dissects distinct lineage trajectories in human myoblast differentiation.

(a) Signalling entropy (SR) versus time point (0 h, 24 h, 48 h, 72 h) for a total of 372 single cells, collected during a time-course differentiation experiment of human myoblasts (scRNA-Seq from Trapnell et al.). Violin plots show the density distribution of SR values at each time point. P value is from a one-tailed Wilcox rank-sum test comparing timepoint 0 h to 24 h. (b) SCENT Gaussian Model fit to SR values predicts three potency states (PS1, PS2, PS3). (c) Probability distribution of potency states at each timepoint. (d) Co-expression heatmap of highly variable genes obtained by SCENT predicting three main clusters. Single cells have been ordered, first by cluster, then by potency state and finally by their time of sampling, as indicated. Landmarks are indicated by rectangular boxes, and distribution of single cells across landmarks and time points is provided in table. Genes have been clustered using hierarchical clustering. Genes that are markers of the different landmarks have been highlighted. (e) Inferred lineage trajectories between landmarks. Diagram illustrates an inferred two-phase trajectory, with one trajectory describing myoblasts of high potency (t=0, cyan circle) differentiating into skeletal muscle cells of intermediate potency (t=24 and 48) (blue circles) and a mixture of terminally differentiated and intermediate potency skeletal muscle cells (t=72) (grey and blue circle, respectively). A second trajectory/landmark describes a different cell-type (interstitial mesenchymal cells) whose intermediate potency state does not change during the time-course (blue stars).