Figure 1

Effects of BLT2 antagonist LY255283 on the survival of 253J-BV cells. Treatment with LY255283 induced cell cycle arrest and subsequent apoptotic cell death in the bladder cancer cell lines 253J (B) and 253J-BV (C), but, not SV-HUC-1 cells (A). Cells were starved with 0.5% FBS medium for 12 h, and then cells were treated with the indicated concentration of each antagonist. Cell cycle distribution (A, B and C left panels) was measured by flow cytometry. Cells were treated with LY255283 (10 µM) or U75302 (1 µM) for 42 h. Cell viability (A, B and C right panels) was detected by MTT assay (as described in the Supplemental material and methods) after treatment with LY255283 and U75302 for 48 h. Data indicate the means ± S.D. of three independent experiments (^*P <0.01, ^**P< 0.001). (D) LY255283-induced cell death was visualised using an Olympus BX51 microscope at ×100 magnification. Bar, 100 µm. (E) LY255283 induced PARP cleavage and activation of caspase-9 in 253J-BV cells. The level of pro-apoptotic protein Bax was increased significantly when cells were treated with LY255283 for 48 h. (F) LY255283 caused internucleosomal DNA fragmentation in 253J-BV cells. The detail apoptotic analysis is described in the Supplemental material and methods.