Fig. 4.

pEZH2(T367) is promotes breast cancer cell migration, invasion, and adhesion without affecting cell proliferation. a Western blot analysis of MDA-MB-231 breast cancer cells showing EZH2 knockdown after lentiviral transduction with control shRNA (shVector) or 3′ UTR EZH2-targeting shRNA (shEZH2) and rescue with myc-tagged WT-EZH2 or T367A-EZH2. b Cells described in a employed in a high-throughput microfluidic migration platform. Data colleted from four independent biological replicates. Box graphs were plotted using Origin 9.0. The bottom and top of the box are the first and third quartiles, and the band inside the box is always the second quartile (the median). The ends of the whiskers represent the 5th percentile and the 95th percentile. The square inside the box indicates the mean, and the x outside the box indicates the minimum and maximum of all of the data. c Cells described in a were seeded in 12-well plates, grown to confluence, and subjected to wound healing assays. Representative images of the wound after 24 h shown above bars. d Reconstituted Boyden basement membrane invasive chamber assay of cells in a. Representative chambers after crystal violet staining shown above bars. e Cells described in a employed in a cell attachment assay. f Cells described in a subjected to time course proliferation assay using Hoescht 33258 to quantify dsDNA. Data for c–f are from at least three independent experiments carried out in at least triplicate and are presented as mean±SD. *p ≤ 0.05; **p ≤ 0.01; ***p ≤ 0.005; ****p ≤ 0.0001