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. 2015 Sep 23;6(33):34758–34773. doi: 10.18632/oncotarget.5325

Figure 2. Breast TAMs and IL-4-activated MDMs promoted the migration of HUVECs via CCL18.

Figure 2

A. Boyden chamber migration-assay results for HUVECs seeded into upper cell-culture inserts, with culture medium alone (Med), medium with IL-4 (IL-4), unactivated MDMs (Ua), IL-4-activated MDMs (M2), MDMs previously cocultured with MDA-MB-231 cells (MDMs-231) or primary breast cancer cells (MDMs-PTu), or primary breast TAMs (TAM) in the lower chambers. B. Boyden chamber migration-assay results for HUVECs seeded into upper cell culture inserts, with TAMs (TAM) plated in the lower chambers in the presence or absence of anti-CCL18 antibodies at 5 (Ab5), 10 (Ab10), or 15 μg/mL (Ab15); anti-VEGF antibodies at 0.5 (Ab0.5), 1.0 (Ab1.0), or 1.5 μg/mL (Ab1.5); combined anti-CCL18 (10 μg/mL) and anti-VEGF (1.0 μg/mL) antibodies (Ab10+Ab1.0); or an isotype-matched IgG control (IgG). C. Boyden chamber migration-assay results for HUVECs seeded into upper cell-culture inserts, with unactivated MDMs (Ua) or IL-4-activated MDMs (M2) plated in the lower chambers, which were untreated (Un), mock-transfected, or transfected with a GFP-siRNA or either of the 2 CCL18-siRNAs. D. Boyden chamber migration-assay results for HUVECs treated with rCCL18 (20 ng/mL), rCCL20 (20 ng/mL), rVEGF (10 ng/mL), or combined rCCL18 (20 ng/mL) and rVEGF (10 ng/mL) (C+V) added to culture medium in the lower chambers. (A–D) Scale bar, 50 μm. Bars correspond to means ± SEMs obtained from 3 independent experiments. ***p < 0.001 versus HUVECs treated with medium alone; ##p < 0.01 and ###p < 0.001 versus HUVECs cocultured with untreated TAMs (B), untreated MDMs (C), or HUVECs treated with rCCL18 (D).