Figure 1 | Scientific Reports

Figure 1

From: Extracellular matrix stiffness dictates Wnt expression through integrin pathway

Figure 1

Wnt/β-catenin pathway was activated by the stiff ECM.

Results from primary chondrocytes 48 hr after seeding on stiff (100 kPa) or soft (0.5–1 kPa) ECM. (a) Microarray profiling of Wnt/β-catenin pathway transcripts. Results are normalized by median scaling using Rosetta Resolver System software. (b) Wnt1 and Wnt3a levels were analyzed by western blotting. (c) Total and phosphorylated ERK1/2 levels were analyzed by western blotting. (d) Axin2, CD44 and (e) phosphorylated GSK3β levels were analyzed by western blotting. (f) Total and phosphorylated β−catenin levels were analyzed by western blotting. (g) β−catenin levels in nucleus and cytoplasm were analyzed by western blotting. (h) Total and (i) activated β-catenin levels and distribution in chondrocytes 2 hr after seeding on stiff or soft ECM were analyzed by in situ fluorescence staining. (j) β-catenin and wnt1 levels in chondrocytes 48 hr after seeding on the Matrigel-coated PAAM were analyzed by western blotting. (k) β-catenin and wnt1 levels in chondrocytes 48 hr after seeding on the ColII-coated PAAM were analyzed by western blotting. Western results were from 3 independent experiments for each individual protein, with blots exemplifying one experiment and the bar graphs showing the combined results of 3 experiments on stiff matrix expressed as percentages (mean ± SEM) of the corresponding results on the soft matrix. GAPDH was used to normalize for equal loading. *P < 0.05, **P < 0.01. n.s. stands for not statistically significant.

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