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Fig. 3 scRNA-seq resolves osteoblast trajectories in the skull. A Lineage analysis and (B) pseudotime analysis of reclustered osteogenic subset using Monocle 3. The black lines in (A) represent the lineage relationship between cells along the trajectory. In (B), cells at the beginning of the trajectory are darkest (purple) and more mature cells are progressively lighter (orange and yellow). C Expression plots of select genes across pseudotime. D Violin plot split by age for osteogenic markers from Monocle analysis. E, F In situ analysis of juvenile frontal (F) bone and adult coronal sutures for mmp9, spp1, and ifitm5. Dotted lines outline the frontal and parietal (P) bones. Arrows mark enriched signal at bone tip and suture edges (mmp9) or along bone surfaces (spp1, ifitm5). In situs were performed in biological triplicate. G Orthogonal section of adult coronal suture labeled by mmp9:GFP and sp7:mCherry. Arrows mark edges of the coronal suture. N = 2. Model of osteogenic cell types at bone fronts and cranial sutures. Scalebars: 100 μM.