Fig. 2.

The soft callus disappears earlier in DIO mice as compared to lean mice. (A) Representative images of IF staining of Col I (red) and Col II (green) in callus tissues isolated from either lean or DIO mice at the indicated post-fracture time points. DAPI stains nuclei (blue). Scale bar = 200μm. (B) Quantitation of Col II-stained area in IF images. Col II-stained area was normalized to the total-callus area and presented as area (%). (C) Quantitation of the mean intensity of Col II staining in IF images. Analysis was performed in 6 lean and 6 DIO callus tissues; 3 sections were analyzed in each. (D) RT-qPCR quantitation of Col2a1 mRNA (normalized to the level of β-actin mRNA). Quantitation was done on RNA purified from lean and DIO callus tissues at d14 post-fracture (n=3). Normalized level in lean callus is defined as 100.

All bar graphs represent average ± SEM. (*) P < 0.05 using student t-test and time-matched lean callus as a control.