Figure 4

Effect of LIMK1 expression on Taxol treatment. A, C: Upper panel: Two-parameter histogram of cell cycle profile of asynchronous BPHLacZ (A) and BPHL (C) cells. Lower panel: BrDU incorporation by asynchronous BPHLacZ and BPHL cells. B, D. Upper panel: Distribution of BPHLacZ cells (B) and BPHL cells (D) in different phases of the cell cycle following Taxol-treatment for 24 h showing accumulation of BPHLacZ cells but not BPHL cells in G2/M containing 4N DNA. Lower panels show limited incorporation of BrDU in both cell types. E. Percentage distribution of BPHLacZ and BPHL cells in G1/S and G2/M phases with or without treatment with Taxol. F. Microscopic analysis of morphology of BPHLacZ and BPHL cells following Taxol treatment (scale bar: 5 μM). Arrows indicate multinucleated giant BPHLacZ cells as noted by DAPI staining. G: quantitative analysis of multinucleated cells, cells in metaphase and cells in anaphase/telophase following Taxol treatment of BPHLacZ and BPHL cells. Five hundred cells were counted for percentage analysis. Data represents the profile obtained from two separate experiments. H. MTT assays of the metabolic activity of cells treated with Taxol or DMSO. Equal amounts of cells were seeded and harvested at 24 and 48 h. Data shows a significant reduction in metabolic activity representing inhibition of cell proliferation in BPHLacZ cells following Taxol treatments. BPHL cells showed proportionately increased metabolic activity with or without Taxol treatment. Data represents Mean+ SD of three separate experiments (* P < 0.001 compared to BPHLacZ DMSO 24 h; **P < 0.02 compared to BPHLacZ DMSO 48 h; • p < 0.01 compared to BPHLacZ Taxol 24 h; •• P < 0.05 compared to BPHLacZ Taxol 48 h). Untreated BPHL cells showed a slower rate of growth compared to BPHLacZ cells.