Fig. 3.

TAM derived IL-6 enriches CSCs in breast cancer. A, B 4T1 cells were treated with CM of activated RAW264.7 cells for 24 h and ALDH1 activity and expression of Sca-1 were examined by FACSCanto II analyser. C 4T1 cells were treated with CM of activated RAW264.7 cells for 24 h and expressions of Sox-2, Oct 3/4 and Nanog were analysed by western blot using their specific antibodies. Actin was used as loading control. Densitometry analysis was performed using NIH-ImageJ software. Relative fold changes with respect to control are shown. D 4T1 cells were treated with CM of activated RAW264.7 cells and seeded in low attachment plates for mammosphere formation. E Bar graph represents fold change in number of mammospheres vs control, mean ± SEM, *denotes p < 0.05, n = 3 independent experiments. F, G 4T1 cells were treated with either CM of activated RAW or CM of activated RAW neutralized with IL-6 antibody (20 µg/ml) and ALDH1 activity as well as Sca1 expression were examined by flow cytometry