Figure 3: Lsd1 restrains migration and invasion of TSCs.
(a) Representative images of TSC colonies showing the morphology of WT, Lsd1 iKO, Ctrl iKD and Lsd1 iKD TSCs after administration of Tx or Dox for 8 days in stemness conditions in comparison to WT TSCs after 2 days of differentiation. Scale bars represent 100 μm. (b) FACS analyses of cell size of wild-type TSCs on day 0 (green), day 2 (blue) and Lsd1−/− TSCs (red). (c) Migration assays of Lsd1 iKO TSCs in the absence or presence of Tx under stemness or differentiation (Diff) conditions recorded in real time. Depletion of Lsd1 in iKO cells is controlled using western blot. Tubulin is used as a loading control. (d) 3D-matrix invasion assays of Lsd1 iKO TSCs in the absence or presence of Tx under stemness or differentiation (Diff) conditions recorded in real time. Cells that invaded the matrix and reached the lower surface of the transwell filter are stained by crystal violet. Scale bars represent 3 mm. (e) Migration assays of undifferentiated Lsd1 iKO TSCs transfected with an Lsd1-expression plasmid (Lsd1) or an empty vector (Ctrl) in the absence or presence of Tx demonstrate that Lsd1 expression rescues increased migration of Lsd1 iKO cells. Bars represent mean±s.e.m. or +s.e.m. Experiments (c–e) were independently repeated at least three times in triplicate. Statistical analysis was performed using two-tailed Student’s t-test. **P<0.01; ***P<0.001.
