Fig 1: FUT11 was up-regulated in PC and predicted poor outcome. (A) qRT-PCR analysis on the mRNA expression of ADM, C4ORF3, ERO1L, FUT11 BNIP3L, KDM3A, MXI1, NDRG1 and KCTD11 in AsPC-1 and PANC-1 cells under normoxia and hypoxia. (B) qRT-PCR analysis of mRNA level of FUT11 in 62 pairs of matched PC tissue and adjacent normal tissues. (C) Western blot analysis of protein level of FUT11 in PC tissue and adjacent normal tissues. (D) IHC analysis of the protein level of FUT11 in pancreatic cancer and normal pancreatic tissue. (E) 90 patients with PC were divided into high- and low-expression groups based on the expression of FUT11. Kaplan survival curve showed the overall survival of high FUT11(red) and low FUT11(blue) expression group. *P < 0.05; **P < 0.01; ***P < 0.001.
Fig 2: Overexpression of PDK1 reversed the inhibitory effect of FUT11 inhibition on PC cell proliferation and migration. Cells were divided into four groups: negative control group (NC), PDK1 overexpressed group (PDK1-UP), FUT11 knockdown group (FUT11-KD) and PDK1 overexpressed plus FUT11 knockdown group (PDK1-UP + FUT11-KD). All cells were cultured in hypoxia. (A) CCK-8 assay on cell viability of each group. (B) EDU assay on cell proliferation of each group. (C) Colony formation on the colony forming ability of cells in each group. (D) Transwell assay on the cell migratory ability of each group. (E) Western blot on the protein level of FUT11, PDK1, N-cadherin and E-cadherin in each group. (F) Western blot on the protein level of phosphorylated AKT, AKT, phosphorylated mTOR and mTOR in each group. *P < 0.05; **P < 0.01.
Fig 3: Identification of key hypoxia-related genes in PC. (A) Intersection analysis of differentially expressed genes in PC samples under hypoxia based on the gene expression profiles of GSE67549 and GSE9350. (B) The mRNA expression of ANGPTL4, ANKRD37, ANKZF1, ADM, C4ORF3, ERO1L, FAM162A, FUT11, BNIP3L, KDM3A, MXI1, NDRG1, KCTD11, PDX1, PFKFB1, PPF1A4, SLC2A1 and P4HA1 in PC tissues compared with non-tumor tissues analyzed by GEPIA online tool.
Fig 4: Overexpression of FUT11 reversed the inhibitory effects of HIF1α knockdown on PC cell proliferation and migration under hypoxia. PANC-1 and AsPC-1 cells were divided into four groups: negative control (NC); FUT11 overexpression (FUT11-UP); HIF1α inhibition (si-HIF1α); HIF1α inhibition plus FUT11 overexpression (si-HIF1α+FUT11-UP). All groups of cells were cultured in hypoxia. (A) Western blot on the expression of HIF1α and FUT11 in each group of cells. (B) CCK-8 assay on the cell viability in each group of cells. (C) EDU assay on the proliferation in each group of cells. (D) Colony formation assay on the colony forming ability in each group of cells. (E) Transwell assays on the migratory ability in each group of cells. The data were shown as means ± S.D. of three independent assays. *P < 0.05, **P < 0.01.
Fig 5: FUT11 bound to PDK1 and regulated its expression. (A) Immunoprecipitation on the binding between FUT11 and PDK1. (B) Immunofluorescence showed FUT11 co-localized with PDK1 in PC cells.(C) Western blot on the expression of FUT11 in sh-scramble and FUT11 knockdown PC cells under normoxia and hypoxia. (D) The ubiquitination assay showed the ubiquitination level of PDK1 in sh-scramble and FUT11 knockdown PC cells under normoxia and hypoxia. (E) CHX was used to inhibit the protein synthesis, and the degradation of PDK1 in sh-scramble and FUT11 knockdown cells under hypoxia detected using Western blot. (F) Western blot on the expression of PDK1 in sh-scramble and FUT11 cells treated with MG132 under hypoxia. (G) IHC images showed the co-expression of FUT11 and PDK1.
Supplier Page from Abcam for Anti-FUT11 antibody