Fig. 1.

OxLDL stimulates classic NF-κB activation through FAK. (A) IKKβ kinase activity (Kin. Act.) in HAE cells treated with oxLDL (100 μg/ml) for the indicated times or with TNFα (1 ng/ml) for 10 min. Beads plus lysate (B+L) served as a negative control and IKKβ activity is expressed as the mean±s.e.m. fold change (Δ) compared to unstimulated conditions (n=4). (B–E) Endothelial cells were transfected with IKKβ siRNA and oxLDL-induced (B) NF-κB phosphorylation (P-NF-κB; 60–120 min, immunoblotting), (C,D) nuclear translocation (60 min, p65 immunocytochemistry), or (E) VCAM-1 expression (exp., 6 h, immunoblotting) was determined. For nuclear translocation, at least 100 cells were scored for NF-κB positivity per condition for each experiment (n=4). (F,G) FAK signaling in HAE cells was inhibited with (F) PF-573228 (4 μM, 1 h) or (G) FAK siRNA and oxLDL-induced IKKβ kinase activity was assessed (n=4). Results are mean±s.e.m. *P<0.05 (compared to 0 time or no treatment) and ^#P<0.05 (compared to respective timepoint) using either (A) one-way ANOVA or (B–G) two-way ANOVA with Bonferroni post-hoc test. NT, no treatment. Mock indicates the mock transfection.