FIGURE 9.

SNX27 is required for HeLa and mpkCCD cell migration. a, HeLa (i) and mpkCCD (ii) cells were infected with a panel of lentiviral SNX27 or control shRNA and selected with puromycin. shRNA expression was induced with 1 μg/ml of doxycycline for 72 h. At this time point, the cells were lysed and expression of SNX27 assessed by Western blotting (top). Equal loading was confirmed by Western blotting using an anti-α-tubulin antibody. b, control and SNX27 knockdown (shRNA#5) cells were cultured to confluence, the wound was created with pipette tip scraping and photographed at the indicated time points. Photographs were taken at 594 nm to mark shRNA expressing cells. Data shown are representative of three independent experiments. Six HeLa (b, i) or three mpkCCD (b, ii) cells per experiment at the edge of the scraped wound were individually tracked and the velocity (μm/s) of each was calculated over at least a 12-h period. The histograms represent the mean velocity; the error bars represent ± S.E. (in μm/s) of all cells measured. SNX27 knockdown cells migrate significantly slower than controls (*, p < 0.0001; **, p < 0.003; two-tailed t test).