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. 2014 Jan 30;289(11):7897–7906. doi: 10.1074/jbc.M113.509497

FIGURE 4.

FIGURE 4.

WASP is required for migration in dense ECMs and infiltration into tumor cell spheroids. BMMs from WT (WASp+/+) and WASP-deficient mice (WASp−/−) were seeded on the top of a thick layer of Matrigel (A; a poorly porous matrix) or fibrillar collagen I (B; a porous matrix), and the percentage of three-dimensional migrating cells was quantified after 48 h. Results are expressed as mean ± S.D. from n = 6 (WASp+/+) and n = 9 (WASp−/−) from three independent experiments. ***, p < 0.001. C, tumor cell spheroids were co-incubated for 3 days with CellTracker-stained BMMs, fixed, and examined with a multiphoton microscope. Spheroid cross-sections are shown. WT macrophages infiltrate the spheroid deeply, whereas WASP−/− macrophages are found outside the spheroid and infiltrated in the peripheral layer. D, quantitation of C, experiments were performed at least in triplicate and expressed as the percentage of macrophages inside spheroids (100% corresponds to macrophages inside plus macrophages adhering on the outer part of the spheroids). Results are expressed as mean ± S.D. (n = 5 WT and 3 WASP−/−; ***, p < 0.001). 3D, three-dimensional.