Effect of stromal fibroblast ECM on the expression of integrins β1 and α2, and phosphorylation of FAK and Src in HCT-116 cells. (A) HCT-116 cells were cultured for 48 h in the absence of matrix (Ctrl), in the presence of stromal fibroblast ECM (Fibrob. ECM) and then immunostained with anti-integrin-β1, anti-integrin-α2 or anti-phospho-FAK. The staining was analyzed through flow cytometry. (B) The relative staining was determined by densitometric analysis. (C) HCT-116 cells were seeded on stromal fibroblast ECM and cultured for 48 h. Lysate proteins were separated on 10% SDS-PAGE and electro-transferred to PVDF membrane. Membranes were blocked and incubated using anti-FAK, anti-phospho-FAK, anti-Src, anti-phospho-Src and anti-β-actin (loading control). Antibody binding was visualized by chemiluminescence and the relative levels of these proteins were determined by densitometric analysis (D). *p ≤ 0.05 compared to control.
Vicente et al. BMC Cell Biology 2013 14:25 doi:10.1186/1471-2121-14-25