Figure 1

Target engagement in CLL cells. (A) Purified CLL cells were incubated with increasing doses of antagonist or vehicle control for 2 h. Subsequently, the synthetic PPARα agonist GW590735 (purchased from GlaxoSmithKline) was added at 1 µmol/L for 48 h. Cells were harvested for RNA isolation. PDK4 (a PPARα target gene) expression was measured by real-time PCR. Data are mean ± standard error of the mean (SEM) from six independent experiments using six different CLL donors. Significant difference: *p < 0.05, unpaired Student ttest. (B) Purified CLL cells were preincubated with vehicle control (veh), NXT629 or the control compound NXT962, both at 30 µmol/L for 2 h. Subsequently, GW590735 was added at 1 µmol/L for 48 h. Cells were analyzed as above. One representative result of two independent experiments using two different CLL donors is shown. Significant difference: *p < 0.05, **p < 0.005, unpaired Student t test. n.s., Nonsignificant. (C, D) Purified CLL cells were incubated in serum- and glucose-free RPMI with NXT629 for 2 h. Subsequently, the natural agonist OEA was added at 10 µmol/L for an additional 4 h. Cells were analyzed for PDK4 (C) or CPT1A (D) as above. Data are the mean ± standard deviation (SD) from independent experiments using four different CLL donors. Significant difference: *p < 0.05, unpaired Student t test.