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. 2013 Jan 1;126(1):312–326. doi: 10.1242/jcs.116244

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© 2013. Published by The Company of Biologists Ltd

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Fig. 3. — Villin regulates directionally persistent cell migration in confluent epithelial monolayers. (A) Directional migration of MDCK cells expressing SEYFP (a), SEYFP-tagged VIL/WT (b) or SEYFP-tagged VIL/Δ21–67/112–119 (c) was determined by tracing the path of migration of cells present at the wound edge over a period of 7 h using the cell track function in Metamorph software. The velocity of cell migration was calculated and compared for 30 individual cells. Values are the measured means ± s.e.m. Cell migration directionality was quantified by measuring as a function of distance along the trajectory L, and the average tortuosity as a function of time t, that were scored for 30 individual cells. The error bars indicate 95% confidence intervals. (B) The directional migration of MDCK VIL/NULL cells and cells expressing VIL/WT or VIL/Δ21–67/112–119 was measured by determining the orientation of the centrosome compared to the cell boundary. Centrosomes were identified by staining for pericentrin. Propidium iodide was used for nuclear staining (a), and the phase-contrast images (b) were merged (c) to determine the leading edge of motile cells. The percentage of cells with the centrosome oriented towards the leading edge was quantified. Values are means ± s.e.m. (n = 15). (C) Directional cell migration in the presence of external stimuli was measured using a modified Boyden chamber. VIL/NULL cells (a) were compared with VIL/WT (b) and VIL/Δ21–67/112–119 (c) cells. Values are means ± s.e.m. (n = 15).

Inline graphic — Villin regulates directionally persistent cell migration in confluent epithelial monolayers. (A) Directional migration of MDCK cells expressing SEYFP (a), SEYFP-tagged VIL/WT (b) or SEYFP-tagged VIL/Δ21–67/112–119 (c) was determined by tracing the path of migration of cells present at the wound edge over a period of 7 h using the cell track function in Metamorph software. The velocity of cell migration was calculated and compared for 30 individual cells. Values are the measured means ± s.e.m. Cell migration directionality was quantified by measuring as a function of distance along the trajectory L, and the average tortuosity as a function of time t, that were scored for 30 individual cells. The error bars indicate 95% confidence intervals. (B) The directional migration of MDCK VIL/NULL cells and cells expressing VIL/WT or VIL/Δ21–67/112–119 was measured by determining the orientation of the centrosome compared to the cell boundary. Centrosomes were identified by staining for pericentrin. Propidium iodide was used for nuclear staining (a), and the phase-contrast images (b) were merged (c) to determine the leading edge of motile cells. The percentage of cells with the centrosome oriented towards the leading edge was quantified. Values are means ± s.e.m. (n = 15). (C) Directional cell migration in the presence of external stimuli was measured using a modified Boyden chamber. VIL/NULL cells (a) were compared with VIL/WT (b) and VIL/Δ21–67/112–119 (c) cells. Values are means ± s.e.m. (n = 15).