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Fig. 2 | Breast Cancer Research

Fig. 2

From: Expression of ID4 protein in breast cancer cells induces reprogramming of tumour-associated macrophages

Fig. 2

Inhibitor of differentiation 4 (ID4) expression in breast cancer cells enhances macrophage motility. a Control of differentiation markers by fluorescence-activated cell sorting analysis in mouse bone marrow-derived macrophages before (T0) and after (T6) culturing in CSF1-rich medium (L929) for 6 days. b Efficiency of ID4 depletion in the SKBR3 cells used for migration assays, evaluated by Western blotting. c Migratory capacity of mouse bone marrow-derived macrophages in response to SKBR3 breast cancer cells, depleted (si-ID4) or not depleted (si-SCR) of ID4 expression, evaluated by Transwell assay. d Efficiency of hemagglutinin (HA)-tagged ID4 overexpression (ID4-HA) compared with that of empty vector transfection (EV) evaluated by using an anti-HA antibody in Western blot analysis. ID4-HA and EV MDA-MB-468 cells were used to prepare conditioned media (CM) for in vivo Matrigel assay. e Schematic representation of Matrigel assay. f and g IHC analysis of mouse macrophage marker F4/80 on Matrigel plugs containing the indicated CM and recovered from mouse flanks at day 7 after inoculation. Counts of F4/80+ cells are indicated in (g). Results from at least three biological replicates are shown. Data are presented as mean ± SEM. ***P < 0.0005 calculated by two-tailed t test

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