Fig. 3

Cytokine chip analysis delineates a different cytokine production signature of CTCs. a The cytokine profile array from the cell culture supernatants of the CTC-TJH-01, 95-D and A549 cells were detected by capture antibodies spotted in duplicate on nitrocellulose membranes. b The quantitative analysis of cytokine profile in A. c The fold changes of CX3CL1 and CXCL5 in the CTC-TJH-01, 95-D and A549 cells. d Verification of CX3CL1 and CXCL5 expression in the CTC-TJH-01, 95-D and A549 cells by western blot. e Real-time PCR and f western blotting were conducted to examine the mRNA and protein expression of CXCL5 in the CTC-TJH-01 cells transfected with the CXCL5-specific siRNA or a non-specific siRNA as the negative control (NC), respectively. g CCK-8 assay and h cell apoptosis assay were used to determine cell proliferation and apoptosis. i, j A transwell assay was used to determine cell migration and invasion. k ELISA assay was used to determine the CX3CL1 secretion level of cell supernatant. l, m After co-culture of CTC-TJH-01, 95-D and A549 cells with lymphocytes; flow cytometry was used to detect lymphocyte recruitment. Each bar represents the mean ± SD of three separate experiments. *P < 0.05; **P < 0.01; ***P < 0.001