Figure 3
Monocytes were suspended at 2.5 × 106 cells/ml and then transfected with sense or antisense ODNs in serum-free media for 4 hours. Transfection efficiency for all genes was routinely > 80%, as determined by counting fluorescein isothiocyanate (FITC)-transfected cells with fluorescence microscopy and comparing with a DAPI label in the same cells (data not shown). Transfected cells were added to Boyden chemotaxis chambers to determine their migratory activity toward rhuIL-18 (2.5 nM). As shown, monocytes transfected with either antisense p38 or ERK½ showed significant reductions in chemotaxic activity toward rhuIL-18 compared with sense transfected cells (n = number of experimental repeats from independent PB monocyte donors).