Preshearing of ECs at HSS, but not LSS, inhibits SMC-induced EC E-selectin expression. ECs were kept as controls (EC/∅) or cocultured with SMCs in an adjacent-bilayer model (EC/SMC) (A,C,E-G) or a media-separation model (EC/M/SMC) (B,D) for 4 hours (E,F), 24 hours (G), or the times indicated (C,D). The E-selectin mRNA (C-F) and surface protein (G) expressions of these ECs were determined by using Northern blot and flow cytometric analyses, respectively. In some experiments, ECs were presheared at HSS (12 dynes/cm²) for 4 hours (HS4) or 24 hours (HS24) or LSS (0.5 dynes/cm²) for 24 hours (LS24) before SMC coculture (E-G). Control ECs were cocultured with SMCs without preshearing (CL) (E-F). Data are presented as percentage changes in band density from control EC/℘ normalized to GAPDH RNA level (C-F) and are shown as mean ± standard of the mean (SEM) from 3 independent experiments. *P < .05 versus control EC/∅. ^#P < .05 versus control EC/SMC. Results of flow cytometric analysis (G) are representative of triplicate experiments with similar results. ECs incubated with FITC-conjugated control IgG or FITC-conjugated antibody alone were used as IgG or negative controls (ie, Blanks: B). Numbers are mean ± SEM of mean fluorescent intensity for all experiments determined by comparison with corresponding negative controls.