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Fig. 6 | Cell Communication and Signaling

Fig. 6

From: FAK displacement from focal adhesions: a promising strategy to target processes implicated in cancer progression and metastasis

Fig. 6

LD2-LD3-LD4 expression inhibits tumor cell spreading, migration and invasion in a dose-dependent manner a) Expression of GFP LD2-LD3-LD4 leads to dose-dependent defects in Hela cell spreading. Spread area is reduced by 38.4 and 52.2% in low-and high-expressing cells respectively (49.16 ± 0.97 μm, n = 216 control cells, compared to 30.27 ± 0.92 μm, n = 102 low-expressing cells and 23.52 ± 1.28 μm, n = 67 high-expressing cells). Cell spreading was calculated as a function of the diameter of the attached area on fibronectin coated coverslips, 1 h after seeding. b) Expression of GFP LD2-LD3-LD4 leads to dose-dependent defects in migration of HeLa. The rate of migration is reduced by 63 and 83% in low and high-expressing cells respectively, following 16 h of recording (100 ± 1.32, n = 549 in control, compared to 36.62 ± 0.28, n = 263 in low-expressing and 17.03 ± 0.2, n = 227 in high-expressing cells). c) Expression of GFP LD2-LD3-LD4 leads to a dose-dependent reduction in the capacity of MDA-MB231 cells to invade Matrigel, as indicated by quantification, using Hoechst staining to detect all cells and GFP to determine expressors. Invasion efficiency is reduced by 63 and 92% in low-and high-expressing cells respectively, following 72 h of incubation (27.95 ± 4.5%, n = 22,718 control, compared to 10.39 ± 03.06%, n = 8950 low-expressing and 2.3 ± 1.02%, n = 5930 high-expressing cells). Invasion efficiency is calculated as the percentage of cells traversing a fluorescently delineated boundary from a low- to high-serum concentration gel, in a chemotactic gradient. To discriminate between high and low expressing cells, we initially determined the mean GFP intensity of all expressing cells, and then compared this to GFP intensity of individual cells, so as to classify them as high or low expressors. ***; p < 0.0001, **; p < 0.005

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