Fig. 5

Analysis of expression of cytokines and chemokines in the CM-treated macrophages by RT-PCR. a Relative expression of CCL22 and TNF-α in THP-1 cells alone or treated with CM in presence or absence of Etanercept (E1 and E10 = 1 and 10 μg/ml Etanercept). MT = MDA-MB-231 + THP-1 cells. b, c Expression of CCL22 and TNF-α mRNA when THP-1 cells were treated with varying concentrations of Etanercept either alone or in presence of MDA-MB-231 condition media (CM). Due to statistically significant effect of the carrier vehicle of Etanercept on the expression of cytokines and chemokines, each concentration of drug is compared to the equivalent amount of vehicle as control. MT = MDA-MB-231 + THP-1 cells, MTB1 and MTB10 = MDA-MB-231 + THP-1 + Vehicle buffer at a concentrations equal to 1 and 10 μg/ml, MTE1 and MTE10 = MDA-MB-231 + THP-1+ Etanercept at concentartions 1 and 10 μg/ml. Data represent mean ± sem of three independent experiments. ΔCT are normalized to GAPDH. ^*P < 0.05 ^**P < 0.01, ^***P < 0.0001. d Co-expression network obtained from co-expression analysis of CCL22 in immune navigator online tool. THP-1 was chose as cell line and correlation coefficient > 0.6 was considered significant. e Interaction between cancer cells and macrophages through secretion of co-expressed cytokines. The highly secreted cytokines in MDA-MB-231 were derived from GSE51938. For macrophages, highly correlated genes with correlation coefficient > 0.6 are shown. f, g, h Expression of IL10 in THP-1 cells alone or treated with CM in presence or absence of Etanercept (E1 and E10 = 1 and 10 μg/ml Etanercept). MT = MDA-MB-231 + THP-1 cells, MTB1 and MTB10 = MDA-MB-231 + THP-1 + Vehicle buffer at a concentrations equal to 1 and 10 μg/ml, MTE1 and MTE10 = MDA-MB-231 + THP-1+ Etanercept at concentartions 1 and 10 μg/ml. Data represent mean ± sem of three independent experiments. ΔCT are normalized to GAPDH. **P < 0.01