Figure 6
Effect of TNC on FN-deficient HUVECs. (a) Cell adhesion of FN-deficient (shFN) and control (shCTL) HUVECs to a non-coated (NC) surface, or a surface coated with BSA, TNC (10 µg/ml) or pFN (10 µg/ml) was determined 45 min after cell plating (±S.D., n = 3). (b) Phase contrast images of shCTL or shFN HUVECs pre-incubated with function blocking anti-integrin antibodies (anti-αvβ3 or α5β1) or with FUD peptide (300 nM) then cultured for 17 h on NC or on TNC. Bar = 50 µm. Additional images are shown in Supplementary Information (Figs S8–S10). (c) Cellular localization of integrins αvβ3 and α5 in FN-deficient HUVECs plated for 48 h on the indicated substrates was determined by immunofluorescence staining and confocal microscopy. Arrows indicate focal adhesions. Bar = 10 µm. (d) Sparsely plated cells were followed by time lapse video microscopy for 5 h (5–10h after plating). Tracking of at least 100 cells per condition was performed. Velocity and persistence length of shFN and shCTL cell movement was determined on non-coated or TNC-coated dishes (top) and on TNC-coated dishes (bottom), after pre-incubation with blocking anti-integrin antibodies (anti-αvβ3 and α5β1) or with FUD peptide (±S.D., n = 3).
