Figure 7. US28 induced monocyte-endothelial cell adhesion requires the downstream signaling molecules Gαq and PKC.
Transduced THP-1 cells were treated with 1 μg/ml of doxycycline or vehicle (H2O) for 24 hours to induce US28 expression. Before co-culture, cells were pretreated with inhibitors of Gαq (YM-254890, 100 nM) and PKC (Ro-32-0432, 5 μM), or vehicle for an hour. 5×105 THP-1 cells were then co-cultured with HECV cells for one hour, and unattached cells were washed away with media. Numbers of input cells and cells attaching to HECV cells were determined by flow cytometry. The ratios between cells attaching to HECV cells and input cells were converted into a percentage. The graph represented four independent experiments. Statistical significance was determined by performing unpaired two-tailed Student’s t tests with GraphPad Prism® software. *: p<0.05, **: p<0.01, ***: p<0.0001.