Figure 5: Nck binding to WIP negatively regulates invadopodium maturation. | Scientific Reports

Figure 5: Nck binding to WIP negatively regulates invadopodium maturation.

From: WIP and WICH/WIRE co-ordinately control invadopodium formation and maturation in human breast cancer cell invasion

Figure 5

(a) WB image shows eGFP fusions (asterisks) and GAPDH expression in which cell lysates were examined using anti-GFP and -GAPDH antibodies. (b) Scheme of full length WIP and WIP deletion mutants lacking the binding domains for cortactin (ΔCBD), Nck (ΔNBD) or N-WASP (ΔWBD). (c) MDA-MB-231 cells overexpressing WIP or WIP deletion mutants (eGFP, green) ΔCBD, ΔNBD and ΔWBD were plated on rhodamine-gelatin (red, 5 h), fixed and stained for cortactin (cyan). Bars: 10 μm (insets, 5 μm). (d) Quantification of invadopodium formation and degradation normalized to eGFP-expressing cell control values. (e) Confocal images of shWIRE eGFP-expressing MDA-MB-231 cells (green; control, WIP, ΔCBD, ΔNBD and ΔWBD) and co-stained for cortactin (cyan). Bars: 10 μm (insets, 5 μm). (f) Quantification of relative invadopodium formation (number of cells that form invadopodia) and degradation (number of cells that degrade gelatin) normalized to control values. Data show mean ± SD of at least three independent experiments (N ≥ 70 cells/condition). *p < 0.05, **p<0.01, ***p < 0.001 by Chi-square test.

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