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Fig. 6. G1-phase arrest induced by inhibition of iPLA₂ requires p53. (A) BEL treatment for 10 hours increases the number of HCTp53^+/+ cells but not of HCTp53^-/- cells in G1 phase. Both HCTp53^+/+ and HCTp53^-/- cells were treated with increasing concentrations of BEL for 10 hours. The DNA contents of each sample was analyzed by FACS. Data represent the mean of three experiments. (B) Accumulation of p53 and expression of p21 increases with increasing concentrations of BEL. HCTp53^+/+ and HCTp53^-/- cells were treated with increasing concentrations of BEL for 10 hours. Cell lysates were prepared and analyzed by western blot for p53, p21, p27 and actin. (C) Inhibition of iPLA₂ with increasing concentrations of BEL dramatically inhibits the proliferation of HCTp53^+/+ but only mildly inhibits the proliferation of HCTp53^-/- cells. HCTp53^+/+ cells (blue bars) and HCTp53^-/- cells (red bars) were cultured with increasing concentrations of BEL in 24-well microplates for 28 hours, followed by 6 hours of BrdU labelling. Cell proliferation was then determined on a µQuant microplate reader. Error bars represent the mean ± s.d. of three experiments; ^*P<0.05, ^**P<0.005. (D) Accumulation of p53 and expression of p21 increases with increasing concentrations of BEL in HCTp53^+/+ (left). In HCTp53^-/- cells, by contrast, p21 expression increased only slightly (right). Both HCTp53^+/+ and HCTp53^-/- cells were treated with increasing concentrations of BEL for 28 hours in 24-well microplates. Cell lysates were prepared and analyzed by western blot for p53, p21, p27 and actin.

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