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. Author manuscript; available in PMC: 2020 Jul 6.
Published in final edited form as: Nat Cell Biol. 2020 Jan 6;22(1):97–107. doi: 10.1038/s41556-019-0443-x

Figure 3. p120-dependent cell-cell cooperation and migration persistence of collective invasion of glioma cells.

Figure 3.

a, Morphology (1), number of filamentous cell-contacts between neighboring cells (2) and cell growth (3) in response to stable expression of non-targeting (NT) and p120 shRNA in E-98 and E-468 cells on a culture plate coated with rBM. Data represent 196 to 226 cells (a2) and 12–18 cell areas (650×486 μm) from duplicate wells per condition from three independent shRNA lentiviral transductions. P values, two-tailed Mann-Whitney test. Efficiency, stability of downregulation and validation using independent shRNA probes are shown in Extended Data Fig. 5h–j. b-h, Impact of p120 downregulation on glioma cell network functions. b, c, Morphology and effect of p120 downregulation on calcium wave propagation between neighbor cells radially migrating from spheroids after 48h culture on rBM. Data in (b) show one out of 2 independent shRNA transductions. Data in (c) represent 10–19 spheroids per condition from 2 independent shRNA lentiviral transductions. P values, two-tailed Mann-Whitney test. d, Network kinetics (6- and 12- hours time-points) of E-468 cells expressing NT or p120 shRNA invading from spheroids into 3D astrocyte scaffolds. Arrowheads, non-polar cells deficient of protrusions and intercellular connections. Data represent 3 independent shRNA lentiviral transductions. e-g, Cell migration persistence, measured as confinement ratio (distance start-end point / total length of path) over a period of 10–13 h (e); image-sequence based quantification of the median number of cell connections (f) and cell migration speed (g) on 3D astrocyte scaffolds after p120 downregulation, obtained by single cell tracking. Data represent 10–20 (e), 296–387 (f), 286–980 (g) cells from 6–8 movies each capturing 3 spheroids per condition from 3 independent shRNA transductions. P values, two-tailed Mann-Whitney test. h, Time-dependent cell-cell interactions in moving cells during 10–15h of migration. Scatter dot plots in (a2; e-g) show individual cells and median (red line). Scale bars, 100 μm (a), 50 μm (b, d).