Figure 4.

Release of 16:1 fatty acids by stimulated macrophages. (A) The cells were either unstimulated or stimulated by mannan plus laminarin (0.5 mg/mL each) for 1 h (colored bars) in the absence (no inhibition) or presence of the following inhibitors: 10 µM FKGK18, 5 µM bromoenol lactone (BEL), 2 µM pyrrophenone (pyrr), or 10 µM SKF98625. Afterward, total content of 16:1 fatty acids was measured by GC/MS. The fatty acid release was calculated by subtracting the amount of phospholipid-bound 16:1 in stimulated cells from that in unstimulated cells. Results are shown as mean values ± standard error of the mean. (n = 4). ** p < 0.01, significantly different from stimulated cells in the absence of inhibitors. (B) The cells were treated for 36 h with sense or antisense oligonucleotide, or vehicle (control), and calcium- (Ca²⁺)-independent group VIA phospholipase A₂ (iPLA₂-VIA) mRNA expression was analyzed by qPCR. (C) iPLA₂ in homogenates from control, sense- or antisense-treated cells was determined by in vitro assay. (D) After the oligonucleotide treatments, the cells were stimulated and 16:1 release was determined. Results are shown as mean values ± standard error of the mean. (n = 4). ** p < 0.01, significantly different from control cells.