Fig. 3

Nidogen-1 blocks endothelial-induced migration of cancer cells. a The endogenous expression of nidogen-1 (NID1) protein was determined by western blot analysis in HUVECs kept under confluent or subconfluent growth conditions and in the cancer cell lines SK-BR-3 and PC3. Actin was used as a loading control. b Fibronectin-1 (FN1) expression was evaluated by immunoblotting analysis in SK-BR-3 cells exposed to conditioned medium derived from subconfluent HUVECs for 4 days or with increasing concentrations of recombinant nidogen-1 (0, 0.01, 0.05, 0.2, and 0.5 μg/ml). GAPDH was used as a loading control. c SK-BR-3 cells were treated for 4 days with unconditioned medium or conditioned medium derived from subconfluent HUVECs with or without recombinant human nidogen-1 (rhNID1 1 μg/ml), with or without human biglycan (rhBGN 1 μg/ml) or in combination. After treatment, the migratory capacity of the cells was determined using a modified Boyden chamber migration assay (**p ≤ 0.01, *p ≤ 0.05 versus control by unpaired student’s t-test)