Expression of LIMK1 in BPH-1 cells and analysis of cell morphology. A: Western blot analysis of LIMK1 expression in BPH-1 parental cells, BPHLacZ and clones of BPHL cells showing increased expression of LIMK1 in BPHL cells. Expression of GAPDH was used as the loading control. B. Phalloidin staining of F actin showing appearance of lamellipodia and ruffling membranes with accumulation of actin in BPHL cells (arrows) (3, 4). BPHLacZ cells showed more microspikes but no lamelipodia with actin accunulation (arrowheads) (1, 2) (scale bar: 5 μM). C. Panel 1 demonstrates normal mitotic spindle and centrosome number with proper chromosomal alignment at the metaphase plate in BPHLacZ cells (scale bar: 5 μM). Because of the vertical position of the cell only one centrosome is visible. Panels 2 and 3 show abnormal spindle architecture with multiple centrosomes and altered chromosomal alignment in BPHL cells. Panel 4 demonstrates appearance of DAPI stained micronuclei in BPHL cells suggesting the presence of chromosomal instability (arrows). Panel 5 demonstrates the presence of multiple nuclei and micronuclei (arrows) in a cell. D: Western blot analysis of expression of LIMK1 in BPHL and BPHLacZ cells used for immunofluorescence analysis. Expression of LIMK1 in BPH-1 parental cells was used for comparison.