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. 2017 Mar 29;8(21):34670–34677. doi: 10.18632/oncotarget.16668

Figure 3. Involvement of cytokines in the enhanced metastatic ability of LuM cells.

Figure 3

(A) Strategy for identifying metastasis-associated genes. In the expression array analysis, 246 genes showed increased expression levels in LuM cells compared with parent KYSE150 cells. Subsequently, gene pathway enrichment analysis showed a strong association of the candidate genes with the cytokine-cytokine receptor pathways. (B) Expression levels of candidate genes by qRT-PCR. Error bars are standard deviations of the means of duplicate experiments. (C) Cytokine array analysis of the cell culture supernatant from parent KYSE150 cells or LuM cells. The red box indicates LCN2. Upper left and lower right on both membrane indicate reference spot. Information of each position on array were shown in manufacturer's instructions. (D) LCN2 protein expression in conditioned medium and the whole cell lysate by western blotting in parent KYSE150 cells and LuM cells. (E) LCN2 secretion levels of the supernatant from parent KYSE150 cells and LuM cells by ELISA. Data are represented as mean ± standard deviations. **p < 0.01. (F) IL-6 and IL-1β protein expression in the whole cell lysate by western blotting in parent KYSE150 cells and LuM cells. (G) IL-6 and IL-1β secretion levels of the supernatant from parent KYSE150 cells and LuM cells by ELISA. Error bars are standard deviations of the means of duplicate experiments. Data are represented as mean ± standard deviations. **p < 0.01.