Fig. 4

Effects of activated Hh signaling in GP injury and chondrocyte behaviors. a Flow cytometry analysis of skeletal stem and progenitor cell-surface-marker of control and operated tibia GP cells at 3 dpi. lineage^- cells, CD45^-Ter119⁻CD31⁻ cells. n = 3. b Immunofluorescence for stem cell marker CD73 after injury of the tibia growth plate of wild-type mice at 3 days post-injury. White arrows indicate CD73⁺ cells. SOC, secondary ossification center. RZ, Resting Zone. PZ, Proliferating Zone. Scale bars, 50 μm. c Tibia growth plates with EdU administration shortly before analysis at 1, 3, 7, and 14 days after GP injury. Scale bars, 200 μm. d Imaris quantification of EdU⁺ cells in the growth plate of control and operated tibias at 1, 3, 7, and 14 days after GP injury. n = 3. e Gli1 and Ptch1 mRNA expression were evaluated by real-time PCR after administration of SAG (100 nM) for 24 h in ATDC5 cells. n = 3. f A scratch-wound assay was performed on ATDC5 cells grown to confluency after being treated with SAG (100 nmol·L⁻¹). Images were taken at the start of the experiment (0 h) and 12 hours later (12 h). Scale bars, 100 μm. g Quantification of migration by measuring the width of the cell-free zone at the time of the scratch (0 h) and 12 hours after the scratch. n = 3. h Chondrogenic differentiation of ATDC5 cells after being treated with SAG (10 nmol·L⁻¹). The error bar represents the standard deviation of the mean. ns, no statistical significance, *P < 0.05, ***P < 0.001