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. 2015 May 14;9(8):1626–1635. doi: 10.1016/j.molonc.2015.05.001

Figure 4.

Figure 4

NGF‐mediated neurotrophic effect of breast cancer cells. A) Co‐culture experiments were performed in Transwell Boyden chambers with breast epithelial cells in the upper part and PC12 cells in the lower part. B) Some breast cancer cell lines were able to induce a neurotrophic effect on PC12 cells. Neurite outgrowth was induced in presence of MDA‐MB‐231, 231‐BR, MCF‐7, SKBR‐3, JIMT‐1, but not in presence of BT‐474 and the non‐tumorigenic HME. A negative control (with no breast cancer cells) and a positive control (addition of 50 ng/ml NGF) have been added. The results represent the mean of 3 independent experiments ± SD. *p < 0.05, **p < 0.01, ***p < 0.001 for comparison with control. C) Representative pictures showing the effect of breast cancer cell lines and HME on PC12 cells. Quantifications are presented in panel B. D) Dot‐blot analysis for the detection of NGF in breast cancer cell conditioned media and quantification of NGF signal intensity in different breast epithelial cells. E) Impact of blocking anti‐NGF antibodies on breast cancer cell‐induced neurite outgrowth. MCF‐7 cells were co‐cultured for 72 h with PC12 cells in presence or absence of anti‐NGF blocking antibodies (1 μg/ml). Control was without MCF‐7 cells, and isotype antibodies were also tested. The results represent the mean of 3 independent experiments ± SD. ***p < 0.001. F) Representative pictures corresponding to the experiment described in panel E.