Fig. 6

The correlations among fibrous tissues accumulation, osteogenesis and YAP nuclear-cytoplasmic shuttling in vivo. A Tissues obtained from patients with nonunion of fractures were harvested and sliced into multiple sections. These sections were subsequently divided into several small unites to enable thorough analysis following immunofluorescent staining. If the α-SMA positive area was over 50%, the area was classified as the high α-SMA expression area, while less than 50% was considered as low expression. Representative immunofluorescent staining images either with high or with low α-SMA⁺ expression were shown. DAPI, OCN and YAP were also stained as indicated in the images. Scale bars, 100 μm. B The nuclear and cytoplasmic YAP intensity in OCN positive cells was calculated using the software Image J. Nuclear YAP positive cells were defined as cells in which the nuclear YAP intensity/cytoplasmic YAP intensity was over 1. Scatter diagram illustrated the correlation between the percentage of α-SMA⁺ areas and the percentage of nuclear YAP⁺ in OCN⁺ cells in A. We ensured accuracy by having at least three different experimenters analyse all images, with the average value of each experiment being shown. C Scatter diagram illustrated the correlation between the percentage of α-SMA⁺ areas and the percentage of OCN⁺ areas in A. We ensured accuracy by having at least three different experimenters analyse all images, with the average value of each experiment being shown. D The nuclear and cytoplasmic YAP intensity in OCN positive cells were calculated using the software Image J. Nuclear YAP positive cells were defined as cells in which the nuclear YAP intensity/cytoplasmic YAP intensity was over 1. Scatter diagram illustrated the correlation between the percentage of OCN⁺ areas and the percentage of nuclear YAP⁺ in OCN⁺ cells in A. We ensured accuracy by having at least three different experimenters analyse all images, with the average value of each experiment being shown