Fig. 3: A neoblast sub-cluster identified as the most potent stem cell for planarian regeneration.

a UMAP demonstrates sub-cell types. b The classical and identified markers for muscle sub-cell types. c The expression pattern of classical marker genes for muscle sub-cell types. d The expression pattern of ndf-1, tpm-1, and rorb at 0 hpa ST data (left) and sbspon during regeneration (right). e The WISH staining of muscle DV markers, atx1l, and six6, at 0 and 12 hpa. Scale bar, 500 μm. f UMAP demonstrates neoblast subpopulations (left) and neoblast sub-cluster (right). g Cell proportions of neoblast subpopulations during regeneration. Source data are provided as a Source Data file. h The identification of marker genes for C7 and C19. i The expression of C19 marker gene, cml-13, in 0 and 12 hpa ST data (left). The WISH staining of cml-13 at 0 and 12 hpa (middle). Scale bar, 500 μm. The expression of cml-13 in scRNA-seq data (right). j Fluorescent in situ hybridization (FISH) showing expressions of C15 markers, osr2, and 2dbd, in 6 hpa samples. Scale bar, 300 and 20 µm for the enlarged field. k The percentage of colocalization of osr2 and 2dbd positive cells. Data were mean ± S.D. and n = 10 animals. Source data are provided as a Source Data file. l Velocity force field showing the average differentiation trajectories (velocity) for cells located in different parts of the UMAP plot. m Pseudo-time of neoblast cells inferred by Monocle2. n Three clusters of branch-dependent genes identified by BEAM. o Bright-field image showing control and osr2 RNAi planarians at 5 dpa, with 6 days post sublethal radiation. Scale bar, 500 μm. Bottom left number, planarians with the phenotype of total tested. p The percentage of regenerated blastema area to the whole body of the control and osr2 RNAi planarians. Error bars represent standard deviation. Data were mean ± S.D. and n = 5 animals in each group. The p values were determined using a two-sided unpaired Student’s t-test (right). Source data are provided as a Source Data file. See also Supplementary Figs. 6, 7.