Fig. 3
The effect of BMP-12 treatment on tendon associated proteins expression in hASCs. a, b Immunocytochemistry (ICC). The expression of collagen I (a) and collagen III (b) (were stained in red) in hASCs cultured for 7 days in standard medium (control, upper panel), or in medium supplemented with 100 ng/ml BMP-12 (lower panel). Nuclei were stained with DAPI (blue). All pictures present cells from the same donor and the same experiment. Scale bars - 20 μm. c Quantification of ICC results. The collagen I and III area per cell was calculated [μm2] after 5 days culture in medium with or without BMP-12 (50 or 100 ng/ml). Data obtained from 2 donors. For each treatment/protein/donor at least 900 cells were subjected to analysis. Kruskal–Wallis one-way analysis of variance detected no statistical differences between groups (ns). d Western blot analysis for collagen I, collagen III and mohawk. Expression of actin was used as a loading control. Graphs below representative bands demonstrate densitometric analysis (normalized to IOD of corresponding actin). The results presented as means ± SEM from three donors (at least three separate electrophoreses per donor performed and analyzed). Data analyzed using non-parametric Wilcoxon test in comparison to internal control (BMP-12 untreated cells from the same donor and electrophoresis).* - p < 0.05