Fig. 3

AMAP1 is required for the invasive activity of MMTV-PyMT tumor cells. a PyMT tumor cells were transduced with the indicated shRNAs and were subjected to immunoblot analysis with the indicated antibodies. b PyMT tumor cells were transduced with the indicated shRNAs and were then transferred to a Matrigel chamber and assayed for invasive activity. Cells that invaded through the Matrigel were stained with crystal violet, and the number of cells in three distinct regions in a single chamber was quantified. Data are means ± SE from more than three independent experiments. *P < 0.05 (Student t-test or Dunnett test, as indicated); n.s., not significant. c PyMT tumor cells were transduced with the indicated shRNAs and were subjected to live-cell imaging analysis. Nuclei were stained with Hoechst 33342 (blue). Representative differential interference contrast (DIC) images of these cells are shown. Arrowheads indicate membrane protrusions. Scale bar, 20 μm. d The cells with (+) or without (−) membrane protrusions were quantified and the percentage of these cells is shown. e Velocity of migration obtained by the total track distance of each cell divided by time is indicated. Bars represent means ± SE. **P < 0.01; ****P < 0.0001 (Student t-test or Dunnett test, as indicated)