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. 2023 May 2;299(6):104774. doi: 10.1016/j.jbc.2023.104774

Figure 3.

Figure 3

Regulation of tumor cell migration and invasion.A and B, PC3 cells expressing vector or Parkin were reconstituted with K2-WT or K2-DM and analyzed for single-cell motility by time-lapse videomicroscopy in 2D contour plots (A), and the average speed of cell movements (B, top) and total distance traveled by individual cells (B, bottom) was quantified. Each tracing corresponds to an individual cell. The cutoff velocities for slow (blue)- or fast (purple)-moving cells are indicated. Mean ± SD (n = 34–46). ∗∗∗p < 0.0001. C and D, PC3 cells as in (A) were analyzed for invasion across Matrigel-coated Transwell inserts and DAPI-stained nuclei of invaded cells (C, representative images) were quantified (D). The scale bar represents 200 μm. Mean ± SD (n = 12–14). ∗∗∗p < 0.0001. E, PC3 cells with Doxy-induced Parkin expression were reconstituted with K2-WT or K2-DM, seeded in 3D spheroids in a collagen-containing matrix, and cell invasion was visualized by light microscopy (top) and INSIDIA software image reconstruction (bottom) for quantification of core area (green) and invasive area (red). The scale bar represents 200 μm. F, the conditions are as in (E), and the invasion area of 3D spheroids was quantified (min to max; n = 7–9). For all panels, numbers correspond to p values by one-way ANOVA with Tukey's multiple comparisons test. DAPI, 4′,6-diamidino-2-phenylindole; Doxy, doxycycline.