Fig. 5

IL-6 restores the ability of breast cancer cells to proliferate and invade, which was inhibited by adipocytes overexpressing Grem2. A MTV/TM-011 cells were seeded in a 96-well plate and treated with each adipocyte CM or IL-6 recombinant protein (25 ng/ml) for 72 h, followed by MTS assay. B MTV/TM-011 cells were treated with each adipocyte CM or IL-6 recombinant protein (25 ng/ml), and then 3D invasion analysis was performed. Scale bar = 100 µm. D After differentiation of 3T3-L1-mock or 3T3-L1-Grem2 into adipocytes for 9 days, 3D invasion assay was performed by mixing adipocytes-mock or adipocytes-Grem2 with MTV/TM-011 cells. Scale bar = 100 µm. C and E The relative invasion area was measured using the Image J. F MTV/TM-011 or MDA-MB-231 cells were treated with each adipocyte CM or IL-6 recombinant protein (25 ng/ml) for 72 h. The lysates were immunoblotted with the indicated antibodies. H Each adipocyte and MTV/TM-011 cells were co-cultured for 72 h in a transwell system (upper chamber: adipocytes, bottom: MTV/TM-011) with or without IL-6 recombinant protein (25 ng/ml). The lysates of MTV/TM-011 cells were immunoblotted with the indicated antibodies. G and I The bar graphs represent relative band intensity. Two-way ANOVA. *, p < 0.05; **, p < 0.01; ***, p < 0.001; ns, not significant