FIGURE 1.

PRMT6-deficient cells exhibit impaired cell movement. A, cell lysates from control and PRMT6 siRNA-treated U2OS cells were immunoblotted with α-PRMT6 and α-tubulin as a loading control. The molecular mass markers are shown on the left in kDa. B, U2OS cell migration and invasion assays were performed following PRMT6 siRNA-mediated transient knockdown. Cell migration (black bars) and cell invasion (white bars) were measured using modified Boyden chambers with filters coated with gelatin or with air-dried Matrigel, respectively. Cells that had moved to the lower surface of the filters were fixed, stained with crystal violet, and quantified as described under “Experimental Procedures.” Results are expressed as a percentage of cell movement seen in PRMT6 knockdown cells compared with control cells. Data represent the means ± S.D. of two independent experiments performed in triplicate. Statistically significant differences, as compared with respective control conditions, are indicated by **, p < 0.01 (Student's t test). WT, wild type. C, monolayer of confluent siGFP- or siPRMT6-treated U2OS cells were wounded with pipette tips, and the cells were incubated with fresh culture medium for 48 h as described under “Experimental Procedures.” The wounded areas were photographed at the beginning (0 h) and at the end (24 h) of the assay. Photographs (original magnification, ×25) obtained from a representative experiment are shown. Results are expressed as the percentage of wound healing after 24 h. Data represent the means ± S.D. of two independent experiments performed in triplicate. D, U2OS cell morphology was monitored following PRMT6 siRNA-mediated transient knockdown. siRNA-treated U2OS cells were plated on coverslips, and stress fibers and focal adhesions were visualized by phalloidin staining and α-vinculin antibodies, respectively, followed by fluorescence microscopy as described under “Experimental Procedures.” Scale bar represents 50 μm.