Vibrantdio cell labeling solution

HUVECs were pretreated with Ad-Best-3 or Best-3 siRNA prior to TNFα (10 ng/ml) incubation for another 24 h in 35 mm culture dishes at a density of 2×10⁵ cells/ml. After treatment, THP-1 cells were labeled with 5 µM VibrantDiO Cell-Labeling Solution (Molecular Probes, NY, USA) and added to each culture dish and allowed to adhere for 30 min at 37°C, 5% CO₂. The dishes were gently washed to remove non-adherent cells twice and the adherent cells were visualized using confocal microscopy (LSM710, ZEISS, München, Germany).

THP-1 cells were labeled with VibrantDiO® Cell-Labeling Solution (V-22886, Molecular Probes, Grand Island, NY, USA), placed on a confluent HAEC monolayer (1 × 10⁵ HAECs per well) in a 96-well plate (1 × 10⁶ THP-1 cells per well), and allowed to adhere for 10 min. After non-adherent cells were removed, the fluorescent intensity of adhered and total cells applied to the well was measured using a fluorescence plate reader (VICTOR X2, Perkinelmer, Waltham, MA, USA). The adherent cells were visualized using confocal microscopy (LSM710, ZEISS, Germany). The ratio of adherent to total cells was expressed as ‘adhesion (%)’.