Figure 2.

Lnk is required for efficient cell spreading. A) HAECs were transfected with the control siRNA (siCtl) or the Lnk-specific siRNAs (siLnk1, siLnk2), and silencing of Lnk was determined by qRT-PCR, Lnk mRNA level was normalized to HPRT-1. Histogram is representative of 3 independent experiments done in triplicate (percentage of basal level, mean ± se). B) After siRNA treatment, cells were plated on collagen-coated glass coverslip and allow to spread for 30, 60, 120, 180, and 240 min. Cells were fixed and immunostained for vinculin (green) to determine cell surface area (perimeter, pixel²) using MetaMorph software (means±se; n=40). Histogram is representative of 3 independent experiments. C) HAECs were either NI or infected with control AdGFP (MOI 30) or AdLnk (MOI 10, 20, 30). Cell lysates were analyzed by Western blot with an anti-Lnk antibody. Blots were reprobed with an anti-GAPDH antibody to ensure equal loading. D) After Ad infection, cells were treated as in B. *P < 0.05 vs. control.