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. Author manuscript; available in PMC: 2020 Aug 1.
Published in final edited form as: Cancer Immunol Res. 2019 Dec 12;8(2):167–178. doi: 10.1158/2326-6066.CIR-19-0514

Figure 4. IL-13 and CCL17 modulated the expression of ST6GALNAC1 in colon cancer cells.

Figure 4.

A) Upper panel. Human cytokine arrays were conducted using conditioned media of HT-29 co-cultured with M1(HT-M1) or M2 macrophages (HT-M2) and media of HT-29 or M2 cultured alone. Each sample was performed in duplicate. Lower panel. List summarizing relative intensities determined by Image J of the most increased cytokines. B) HT-29, M1 and M2 macrophages supernatants and conditioned medium of HT-29 co-cultured M1 or M2 were harvested for cytokine and chemokine analysis using the LegendPlex chemokine array. C) IL-13 cytokine expression was determined using ELISA assay. D-E) Real-time PCR and western blotting analyses of ST6GALNAC1 in HT-29 cells alone or co-cultured with M2 macrophages with or without IL-13 neutralizing Ab (D) or CCL17 neutralizing Ab (E). F) ST6GALNAC1 and sTn expression in IL-13- and CCL17-treated HT-29 cells was detected by immunoblotting analysis. Actin was used as loading control. G) The expression of mRNA of ST6GALNAC1 in IL-13- and CCL17-treated cells were detected using real-time PCR. GAPDH was used as housekeeping gene. P values were calculated using one-way (C,G) or two-way (B,D,E) ANOVA with Tukey post-tests for multiple comparisons. Error bars represent the SEM (B-E and G). *p<0.05, **p<0.01, ***p<0.001. All results are representative of three independent experiments.