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. 2019 Sep 19;14(9):e0222580. doi: 10.1371/journal.pone.0222580

Fig 10. Invasive capacity of 231/LM2-4 cells under control and 5-FU+4-HC conditions using a 3D lrECM “on-top” assay.

Fig 10

(Control): Mass structures (A): round morphology (1–3), irregular morphology showing collective cell migration as chains of few cells with smooth borders (Δ9), buds (Δ6) or as disorganized masses (Δ5). Single cell protrusions (Δ12,14,16). Multicellular streaming with no apparent junction contacts (Δ7,8). Dissemination of single tumor cells (pink Δ5,16). Pseudo-Stellate Mass structures (B): multicellular collective protrusive migration with leading cells with invadopodia (Δ2,4,5). Dissemination of single tumor cells (pink Δ1) and collective dissemination (pink Δ7). Contacts between several structures (pink Δ6). Stellate structures (C): several multicellular chains with a collective cell migration pattern of invasion and protrusive leading front with one (Δ9) or multiple leading cells (Δ15) with invadopodia. Multicellular invasive chains with 1–2 cells in diameter (Δ13) or broad masses of cells (Δ18). Secondary branches at the lateral margin of the invasive chains (Δ12). Cell dissemination (pink Δ7,14,17). Contacts between several structures (pink Δ22,24). (1μM 5-FU + 0.01μM 4-HC): Mass structures (A): round morphology (1–3), irregular morphology exhibiting collective cell migration as chains of few cells with smooth borders (Δ5), buds (Δ4), and structures with single-cell protrusions (Δ14,15). Multicellular streaming (Δ7,9). Dissemination of single tumor cells (pink Δ11) and group of cells (pink Δ12). Pseudo-Stellate Mass structures (B): multicellular collective protrusive migration with leading cells with invadopodia (Δ3) or leading buds (Δ9). Dissemination of group of cells (pink Δ17). Stellate structures (C): multicellular chains with a collective cell migration pattern of invasion and protrusive leading front with invadopodia (Δ5,7), or leading buds (Δ10). Multicellular invasive chains with few cells in diameter (Δ13) or broad masses of cells (Δ13,20). Single and collective cell dissemination (pink Δ27 of the latter). Multicellular chains with an uncoordinated arrangement of the component cells (pink Δ11), contacts (pink Δ15,26) mainly fusions (pink Δ24,25,27). The experiment was repeated at least three times with similar 231/LM2-4 invasiveness inhibitory effect by the treatments. The best multicellular structures were obtained with MatrigelTM matrix basement membrane (BD BioSciences, Cat. 354234-Lot 88482) with a high concentration (10mg/ml).