Expression of syndecan-2 in HCT-116 cells. (A) Immunoprecipitation of syndecan-2 from HCT-116 cells in the absence of matrix (Ctrl) or in the presence of stromal fibroblast ECM (Fibrob. ECM). HCT-116 cells were cultured for 72h and exposed to [35S]sulfate for 24 h, and the radioactive proteoglycans were extracted as described in Methods. The proteoglycans from the cells were immunoprecipitated with anti-syndecan-2 antibody and were then applied to the gel. (B) Quantification of the experiment shown in A. (C) HCT-116 cells were seeded on stromal fibroblast ECM and cultured for three days. Lysate proteins were separated on 10% SDS-PAGE and electro-transferred to PVDF membrane. Membranes were blocked and incubated using anti-syndecan-2 (Syn-2) 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