Fig. 2.

NADPH oxidase/ROS are involved in LysoPC-induced COX-2 and IL-6 expression. a HCFs were treated with LysoPC (40 μM) for the indicated time intervals. The generation of ROS and activity of NADPH oxidase were determined using a CM-H₂DCFDA probe (open bars; n = 10) and lucigenin chemiluminescence analysis (black bars; n = 10), respectively. b HCFs were pretreated with either DPI (100 nM) or edaravone (Eda; 100 nM) for 1 h and then incubated with LysoPC for 30 min. The cell lysates were subjected to determine the ROS generation (open bars; n = 5) and NADPH oxidase activity (black bars; n = 7). c HCFs were pretreated with DPI for 1 h and then incubated with LysoPC for 6 h. The levels of COX-2 and GAPDH protein were determined by western blot (n = 6). d, e HCFs were transfected with siRNA of scramble, NOX1, NOX2, NOX4, or NOX5 and then incubated with LysoPC for 6 h. The levels of COX-2, IL-6, and GAPDH mRNA were determined by RT/qPCR (d, n = 5; e, n = 5). f HCFs were pretreated with either DPI (100 nM) or SP600125 (1 μM) for 1 h and then treated with LysoPC for the indicated time intervals. The levels of JNK1/2, phospho-JNK1/2, and GAPDH protein were determined by western blot (n = 5). The densitometry measurements are presented in Supplementary Fig. 2B and C. Data are presented with mean ± SEM and analyzed by one-way ANOVA with Tukey’s post hoc tests. *p < 0.05; ^#p < 0.01