Figure 2
Increase in the formation of matrix and ECM contacts in P25-treated MG-63 cells. A – MG-63 cell monolayers were incubated for 16 hours in DMEM containing 2 μg/ml AlexaFluor488 derivatized plasma fibronectin (green) in the presence of 50 μM S25 (a, b) or P25 (d, e). Cells were subsequently fixed, permeabilized and endogenous fibronectin was visualized by indirect immunofluorescence using the IST-9 antibody specific for cell-derived fibronectin (red). Panels c and f show merged images indicating colocalization of P25-dependent fibronectin polymerization with endogenous fibronectin matrix. B – Cells in suspension were allowed to attach for 6 hours on fibronectin-coated coverslips in DMEM containing soluble AlexaFluor488-fibronectin (20 μg/ml, green), in the presence of 50 μM S25 (a, b) or P25 (d, e). Cells were fixed, permeabilized, and activated β1 integrin was visualized by indirect immunofluorescence using the 9EG7 antibody (red). Panels c and f show merged images indicating colocalization of activated integrins with fibronectin fibers.