Journal: Scientific Reports

Article Title: Extracellular matrix stiffness dictates Wnt expression through integrin pathway

doi: 10.1038/srep20395

Figure Lengend Snippet: 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.

Article Snippet: Microarray analyses were performed using commercial Mouse cDNA Microarray slides (Phalanx Biotech Group; Hsinchu, Taiwan) according to the manufacturer’s instructions.

Techniques: Microarray, Software, Western Blot, In Situ, Fluorescence, Staining