Figure 6

Modulation of Wnt/FZD4 signaling pathways through the interaction of miR-204 with FZD4-miR-SNP (rs713065) variant in NSCLC cells. (A) The top-scored network in Cell Cycle, Gene Expression, and Cellular Function and (B) canonical Wnt/β-Catenin signaling pathway associated with FZD4-miR-SNP (rs713065) and Wnt5A activities in H1299 cells were representatively displayed. H1299 cells were transfected with either FZD4-SNP-3′UTR or FZD4-WT-3′UTR. After incubation for 48 h, the total RNAs were isolated and absolute gene expression profiles were determined by nCounter, with differentially expressed pathways identified through Ingenuity® Pathway Analysis. (C) Modulation of top-scored cellular network and (D) Wnt/β-Catenin canonical pathways through the miR-204 targeted FZD4 expression in 3′UTR containing FZD4-miR-SNP. Key molecules identified through NanoString and IPA analysis (red if is up-regulated in H1299 cells co-transfected with FZD4-SNP/Wnt-5a compared to FZD4-WT/Wnt-5a in the presence or absence of the ectopic expression of miR-204, green if is down-regulated). The intensity of the color indicates expression level or degree of regulation. The top-scored networks and signaling pathways and their associated molecules and cellular functions were summarized in details in Supplementary Tables 1–5)