Fig. 1.

LPC induce COX-2 mRNA and protein expression. EA.hy 926 cells were incubated with 200 μM LPC or PBS in medium containing 10% FBS for the indicated time points, followed by the determination of (A) COX-2 mRNA abundance by qRT-PCR and (C) COX-2 protein by Western blotting. In (B) cells were pre-incubated with actinomycin D (ActD) (800 nM) for 30 min before addition of LPC followed by incubation without or with LPC 16:0 for 1 h and LPC 20:4 for 4 h in the absence or presence of ActD. Subsequently, relative COX-2 mRNA expression was determined by qRT-PCR as detailed in Section 2. In (D) cells were pre-incubated with cycloheximide (+CHX) (100 μM) or vehicle (-CHX) followed by incubation without or with LPC 18:2 in the presence or absence of CHX for 5 h. Subsequently, expression of COX-2 protein was analyzed by Western blotting. Results shown in (A) and (B) are mean ± SD of three independent experiments done in triplicates. In (A) the values of LPC samples were normalized to those of PBS, which were set to 1. Results in (C) and (D) are representative Western blots. *, **, *** indicate significant differences between LPC and PBS treated cells. ^#, ^##, ^### indicate significant differences between LPC treated cells in the presence and absence of applied inhibitors.