Fig. 4.

TRIM65 enhances cell migration through actin and microtubule cytoskeleton remodeling. a Representative immunoblots of EMT markers. b Images and statistics of the number of filopodia and focal adhesion of KM12 cells that were stably transfected with TRIM65 or its control Empty, by using Alexa Fluor 488 phalloidin staining for the actin cytoskeletal fibers observed by fluorescence confocal microscopy. Filopodia/focal adhesion are indicated by red arrow heads in dash yellow box. At least 100 cells for each group were counted. Mean ± S.D, n = 3, **p < 0.01, two-tailed Student’s t-test. Scale bar: 100 µm. c Representative images of the mitotic spindle apparatus analyzed by immunofluorescence microscopy after cells were stained for γ-tubulin (green) and DAPI (blue). Left panel is siCTL with normal spindle and right panel is abnormal spindle (polyspindle) after siTRIM65. Scale bar: 12 μm. d Histogram shows the percentage of abnormal spindles in mitotic cells per experiment (siTRIM65 compared with siCTL, at least 100 cells counted for each cell line). Mean ± S.D.; n = 3, *p < 0.05, **p < 0.01, two-tailed Student’s t-test