Figure 4.

(A) Representative hematoxylin and eosin staining images on callus sections from vehicle- and irisin-treated mice at 28 days post-fracture (scale bar: 0.8 mm). The black squares indicate the area of higher magnification (scale bar: 60 µm). Arrowheads indicate the fibrous tissue. (B) Representative Trap staining images of callus sections from vehicle- and irisin-treated mice at 28 days post-fracture (scale bar: 30 µm). (C) Representative images of osteocalcin immunostaining in callus sections from vehicle- and irisin-treated mice at 28 days post-fracture (scale bar: 30 µm). (D) Quantification of OC N./CA in sections from vehicle- (n = 6) and irisin-treated mice (n = 6) at 28 days post-fracture. (E) Quantification of the percentage of osteocalcin-positive cells (osteocalcin⁺ cell, %) out of all cells of the callus from vehicle-treated mice (n = 6) and irisin-treated mice (n = 6) at 28 days post-fracture. Data are presented as dot-plots with medians, from maximum to minimum, with all data points shown. The Mann–Whitney test was used to compare groups.