2.9. Monocyte Adhesion Assay

The adhesion of THP-1 cells to control siRNA-treated or ADAM15 siRNA-treated HUVECs grown to confluence in 96-well plates was examined. HUVECs were serum-starved overnight. The next day, the cells were treated with TNF-α (10 ng/mL) for 24 h. THP-1 cells were collected and labeled with Calcein AM fluorescent dye (Life Technologies, 5 μM) for 30 min. After being washed twice, 1 × 10⁵ THP-1 cells were added to each well and incubated for 30 min at room temperature. Nonadherent cells were washed away, and the fluorescence was measured using a Synergy HT fluorescence plate reader (BioTek Instruments, Winuski, VT, USA).