Fig. 4.

FUT4 mediates the activation of membrane trafficking and oncogenic pathways. (a) Top 15 positivelyenriched gene sets in genome-wide RNA-seq data of A549_FUT4^high and CL1–0_FUT4 cells versus respective vector controls. Enriched gene set data from Gene Set Enrichment Analysis (GSEA) with a nominal p value less than 0.05 and false discovery rate (FDR) less than 0.25 are presented. (b) GSEA enrichment plots of top 8 positively enriched gene sets in A549_FUT4^high and CL1–0_FUT4 cells, including membrane trafficking, cell cycle, TGFβ signaling, RNA processing, hypoxia, metastasis, EGF, and MAPK signaling pathways. (c) Immunofluorescent imaging analysis of cytoskeleton and cell morphology of A549 lung cancer cells with various levels of FUT4 over-expression (A549_vector, A549_FUT4^med, and A549_FUT4^high). Scale bar: 50 μm (left panel) and 20 μm (right panel). (d) Western blot analyses of EMT marker proteins in A549 lung cancer cells with various levels of FUT4 over-expression. (e) Heatmap of RNA-seq transcriptomes for the leading-edge genes of KEGG_pathway_in_cancer from GSEA analyses of TCGA lung adenocarcinoma. Top sidebars denote the mutation status of EGFR and expression levels of FUT4 grouped with the median value cut-off. p value in (a, b) was calculated by one-way ANOVA with Dunnett's test. * p < 0.05, ** p < 0.01, *** p < 0.001. ns: not significant. Experiments were performed in three biological replicates and presented as mean ± SEM.