Photos of fractured tibia at Time 0 0 (T0), and mice intermitFigure 1. (A) Representative radiological images of fractured tibia at Time (T0), and mice intermittently treated with (B) standard saline (automobile) or (C) irisin at 10 days post-fracture. (D) Representative tently treated with (B) standard saline (vehicle) or (C) irisin at 10 days post-fracture. (D) Representative Safranin O-CD1530 Autophagy staining pictures of callus sections from vehicle- and irisin-treated mice at 10 days Safranin O-staining photos of callus sections from vehicle- and irisin-treated mice at 10 days postpost-fracture (scale bar 0.eight mm). The black squares indicate the locations of greater magnification (scale fracture (scale bar 0.eight mm). The black squares indicate the regions of higher magnification (scale bar: bar: 60). (E) Dot-plot graphs displaying the elevated soft callus region and (F) decreased proteogly60). (E) Dot-plot graphs showing the enhanced soft compared with vehicle-treated mice (n = six). can-rich cartilage matrix in irisin-treated mice (n = six) callus area and (F) decreased proteoglycan-rich cartilage matrix in irisin-treated pictures = callus sections from vehicle- (n = mice (n = six). (G) Repre(G) Representative Trap staining mice (n of six) compared with vehicle-treated six) and irisin-treated (n =sentative Trapdays post-fracture (scale bar: 0.8 mm). The black squares indicate the locations of greater 6) mice at 10 staining photos of callus sections from vehicle- (n = 6) and irisin-treated (n = 6) mice at ten days post-fracture (scale bar: 0.8 mm). The black squares increased number of Trap-positive cells magnification. (H) Dot-plot graph showing the considerably indicate the locations of larger magnification. in the callus ICA-105574 Membrane Transporter/Ion Channel location (OC n. /CA) in irisin-treated mice (n = six) comparedTrap-positive cells in mice (n = (H) Dot-plot graph showing the considerably improved number of with vehicle-treated the callusarea (OC n. /CA) in irisin-treated mice (n = 6) compared with vehicle-treated mice (n = six) (scale bar: 60). Information are presented as dot-plots with medians, from maximum to minimum, with all data points shown. The Mann hitney test was employed to examine groups.Int. J. Mol. Sci. 2021, 221,five ofInt. J. Mol. Sci. 2021, 22,6) (scale bar: 60). Information are presented as dot-plots with medians, from maximum to minimum, with all information points shown. The Mann hitney test was employed to examine groups.5 ofImmunohistochemical staining for COL II in callus sections (Figure 2A) and relative Immunohistochemical staining for COL II in callus sections (Figure 2A) and relative quantification (Figure 2B) showed no important difference amongst the two experimental quantification (Figure 2B) showed no significant difference in between the two experimental groups. On the other hand, the expression of COL X, well-established marker of hypertrophic groups. Nonetheless, the expression of COL X, a a well-established marker of hypertrophic chondrocytes, was enhanced threefold (p 0.0012) within the callus of of irisin-treated mice chondrocytes, was improved threefold (p == 0.0012) inside the callusirisin-treated mice compared with all the the automobile group (Figure 2C,D). Of note, the positivity for the master compared with vehicle group (Figure 2C,D). Of note, the positivity for the master regulator of osteoblast differentiation, RUNX2, was two.2-fold larger larger (p = (Figure E-F), whereas regulator of osteoblast differentiation, RUNX2, was two.2-fold(p = 0.0137) 0.0137) (Figure 2E,F), the positivity for SOX9, the transcription aspect that regulates chondr.
