Figure 2
PPARα antagonist is cytotoxic to CLL cells, even in the presence of the microenvironment. (A) CLL cells were cultured in the presence of the PPARα antagonist NXT629 or DMSO control, added once at the beginning of the culture. CLL cells were harvested after 4 d and stained with DiOC6/PI and analyzed by flow cytometry. The percentage of viable cells as determined by gating on DiOC6 bright and PI-negative cells is shown. Data are mean ± SD for five different experiments with five different CLL donors. The IC50 was calculated using GraphPad Prism software. (B) CLL cells were cultured alone or in the presence of mitomycin C-treated J774 macrophages. NXT629 (10 µmol/L) or DMSO was added once at the beginning of the coculture. Cell viability was measured at d 6 as above. Data are mean ± SD from two independent experiments using two different CLL donors. Significant difference: ***p < 0.0005, unpaired Student t test. CLL cells were cultured in the presence (C) or absence (D) of mitomycin C-treated J774 macrophages. DMSO or fludarabine was added once at the beginning of the coculture. Cell viability was measured at d 6 as above. Data are mean ± SEM from two independent experiments using two different CLL donors. Significant difference: ***p < 0.0005, unpaired Student t test. n.s., Nonsignificant. (E) CLL cells were culture alone or in the presence of preadipocytes (OP9) cells. NXT629 (10 µmol/L) or DMSO was added once at the beginning of the coculture. Cell viability was assessed at d 6 as above. Data are mean ± SEM from two independent experiments using two different CLL donors. Significant difference: **p < 0.005, unpaired Student t test.