Figure 7
From: Extracellular matrix stiffness dictates Wnt expression through integrin pathway
Wnt/β-catenin pathway contributes to the regulation of BMMSC Differentiation by ECM stiffness.
(a) β-catenin and Wnt1 levels in BMMSCs 48 hr after seeding on stiff or soft ECM were analyzed by western blotting. (b) BMMSCs were cultured on stiff or soft ECM for seven days in the presence of 10 μM Cardamonin or DMSO, followed by determination of Runx2, ColI expressions western blotting. (c,d) BMMSCs were cultured on stiff or soft ECM for seven days in the presence of 10 μM Cardamonin or DMSO, followed by determination of MAP2, NFL, Runx2 and ColI expressions by immunocytochemical staining. Scale Bar: 30 μm. (e) BMMSCs were cultured on stiff or soft ECM for seven days in the presence of 10 μM Cardamonin or DMSO, followed by determination of MAP2 and NFL expressions western blotting. (f) BMP2 levels in BMMSCs treated with 10 μM Cardamonin or DMSO for 24 hr were analyzed by western blotting. Results on fluorescence intensities from 6 independent experiments on stiff matrix are expressed as percentages (mean ± SEM) of the corresponding results on the soft matrix. Western results were from 3 independent experiments with blots exemplifying one experiment and the bar graphs showing the combined results of 3 experiments on stiff matrix expressed as percentages of the corresponding results on the soft matrix. GAPDH was used to normalize for equal loading. *P < 0.05, **P < 0.01, ***P < 0.001, n.s. stands for not statistically significant.
