Fig. 2

Reconstruction of the myogenic differentiation trajectory in a pseudotime manner. A Pseudotime analysis of myogenic cells (including Pax3+ progenitors, myogenic progenitors, myoblasts, and myocytes in Fig. 1E) was performed by Monocle 2 and revealed seven different cell states (states 1~7). The distributions of cell states were presented along with pseudotime flows. Each dot is a cell. B Visualization of myogenic differentiation trajectory by cell origins (left) and developmental stages (right). C Visualization of myogenic differentiation trajectory by cell identity. D Violin plots showing feature gene expression in each cell cluster. MYMK, FNDC5, MEF2C, and TNNI1 are muscle development-related genes. MYL9 is a cardiomyocyte-specific marker. MYOD1 and MYOG are skeletal muscle cell-specific markers. E Gene Ontology (GO) analysis of the differentially expressed genes with high levels in myocytes (state 2) and myocytes (state 7). F Visualization of myogenic differentiation trajectory by cell state, with cardiac cells distinguished from differentiating skeletal muscle cells. G Bar plot showing the percentage of cells within each sample assigned to the annotated myogenic cell types. H Immunofluorescence staining for Pax7 and MyoD on somite cross sections of ZZ and DZ at E21 and E28. Scale bar = 100μm