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Figure 2


Fig. 2. Inactivation of iPLA2 by BEL arrests INS-1 cells in G1 phase. (A) Inactivation of iPLA2 inhibits DNA synthesis. Cells were cultured for 24 hours in with or without BEL (12.5 µM) and labelled with BrdU or not. Cells were stained with fluorescein-conjugated anti-BrdU for BrdU and PI for DNA. R2 indicates cells in S phase. (B) BEL prevents cell progression to G2-M transition. INS-1 cells were treated with BEL (12.5 µM) followed by immunofluorescent staining of the mitosis marker phosphohistone H3. Treatment of cells with nocodazole served as a control for G2-M-transition arrest. Cells were fixed and stained for phosphohistone H3 and for iPLA2. Magnification, 40x. (C) Inhibition of iPLA2 in the presence of nocodazole increases the number of cells in G1 phase. INS-1 cells were treated either with nocodazole (0.5 µg/ml) only or with both nocodazole and different concentrations of BEL for 20 hours and 30 hours. Nontreated cells were used as controls. The DNA content of the cells was then analyzed by FACS after PI staining. The percentages of each indicated phase represents the mean of three FACS analyses.





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