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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: Nat Immunol. 2012 Jan 8;13(2):136–143. doi: 10.1038/ni.2205

Figure 2. Netrin-1 inhibits macrophage migration to CCL2 and CCL19, via its receptor UNC5b.

Figure 2

(a) Migration of RAW264.7 cells to CCL2 (100 ng/ml) was measured in the absence or presence of increasing concentrations of recombinant netrin-1 placed in the lower compartment of a Boyden chamber. (b) Real-time measurement of migration of RAW264.7 cells to 250 ng/ml netrin-1, 100 ng/ml CCL2, or both. (c) Migration of RAW264.7 cells pretreated (PT) with 250 ng/ml netrin-1 and exposured to CCL2 (100 ng/ml). (d-e) Migration of mouse pMø to (d) CCL2 (100 ng/ml) or (e) CCL19 (500 ng/ml), in the absence/presence of 250 ng/ml netrin-1. (f) pMø stained with phalloidin to detect polymerized actin after treatment with 100 ng/ml CCL2 with or without 250 ng/ml netrin-1. Arrows indicate membrane ruffles, scale bar 10 μm. (g) Mean cell surface area of pMø in (f). (h). Quantification of actin polymerization by flow cytometric analysis of phallodin staining. (i) Amount of activated Rac1 in pMø treated for 5 min with 100 ng/ml CCL2 with and without 250 ng/ml netrin-1. (j) Immunoblot of phospho- and total FAK in pMø incubated with 100 ng/ml CCL2 with or without 250 ng/ml netrin-1 pretreatment. Data are the mean ± s.d. of triplicate samples in a single experiment and are representative of 3 independent experiments. P < 0.05, **P < 0.01.