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Fig. 3 | Molecular Cancer

Fig. 3

From: PKN2 in colon cancer cells inhibits M2 phenotype polarization of tumor-associated macrophages via regulating DUSP6-Erk1/2 pathway

Fig. 3

PKN2 in colon cancer cells induces the differentiation of monocytes into M2 polarized macrophages in vitro. a HCT116 cells were stably transfected with vector/PKN2-WT or shCTL/ shPKN2-1/ shPKN2-2. The protein expression of PKN2 was detected by western blotting in total protein extracts. Human CD14+ monocytes were separated from peripheral blood and co-cultured with vector/PKN2-WT (b) or shCTL/ shPKN2-1/ shPKN2-2 (c) transfected HCT116 for 4 days, respectively. E:T ratios for colon cancer cells to monocytes were 50:1. Flow cytometry was used to explore surface expression of CD206. *, P < 0.05 versus Vector. (d-f) SW480 cells were treated and co-cultured with CD14+ monocytes as indicated in (a-c). Western blotting was used to evaluate PKN2 expression in SW480 cells. Flow cytometry was used to explore surface expression of CD206 in differentiated macrophages. **, P < 0.01 versus Vector. (g)~(i) HT-29 cells were treated and co-cultured with CD14+ monocytes as indicated in (a)~(c). PKN2 and CD206 expression was analyzed. *, P < 0.05 versus shCTL. (j) CD14+ monocytes were treated as indicated in (b), gene expression of IL1B, TNF, CXCL9, IL6, IL10, ARG1 and CCL22 in monocytes was detected. (k) CD14+ monocytes were treated as indicated in (i), gene expression of IL1B, TNF, CXCL9, IL6, IL10, ARG1and CCL22 in monocytes was detected. *, P < 0.05;**, P < 0.01 versus Vector or shCTL

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