Runx2 and p21 are decreased in Bmf-/- diabetic mice. (A-C) The expression of Runx2 (A), p21 (B) and BMF (C) in aorta tissues from the four groups of mice were examined by immunohistochemistry (IHC). Black arrows indicate the positive expression of Runx2, p21 and BMF (n = 5/group). Scale bar in red represents 50 μm. (D) The protein level of Runx2, BMF and p21 in aorta tissues from the four groups of mice. The intensity of each band in WB was quantified by densitometry, and data were normalized to the β-actin signal (n = 5/group). (E) The expression of BMF-AS1 and BMF in aorta tissues as fold changes relative to WT group (n = 5/group). The data were expressed as mean ± SD, one-way ANOVA for Fig. 5A-E. Quantification of positively stained areas was measured by Image-Pro Plus software (version 6.0). ***p < 0.0005, **p < 0.001, *p < 0.05, compared with WT.##p < 0.001, compared with Bmf-/+ diabetic mice. ns: non-significant. WT: wild type; DM: diabetes mellitus.