Fig 1: The NORAD/FUBP1 interaction results in the upregulation of downstream pro-apoptotic genes.a RNA-seq data of liver cancer from TCGA to classify FUBP1 high- and low-expressed groups. b GSEA for the FUBP1-related pathways in liver cancer. c qRT-PCR analysis for the expression of four FUBP1 downstream targets (TRAIL, NOXA, BIK, and TNFA) by knockdown of FUBP1. d qRT-PCR analysis for the expression of four FUBP1 downstream target genes by the introduction of full-length NORAD or the NORAD-1 fragment, which was not bound to FUBP1. e, f ChIP-qPCR analysis for the FUBP1 (e) and RNA polymerase II (f) occupancies at the promoters of four target genes (TRAIL, NOXA, BIK, and TNFA) after transfecting full-length NORAD or NORAD-1 fragment. The fold enrichment was relative to the input DNA. The results were determined from triplicates, and the error bars represented as the mean ± SD, *P < 0.05, **P < 0.01, ***P < 0.001. TNF tumor necrosis factor, TRAIL TNF-related apoptosis-inducing ligand, NOXA PMAIP1, phorbol-12-myristate-13-acetate-induced protein 1, BIK BCL2 interacting killer, ChIP chromatin immunoprecipitation, Pol II RNA polymerase II.
Fig 2: FUBP1 promotes pancreatic adenocarcinoma cell viability. Viability of (A) PaTu8988 cells transfected with si-NC or si-FUBP1 and (B) SW1990 cells transfected with empty or FUBP1-OE vectors was analyzed using a Cell Counting Kit-8 assay. Data are presented as the mean ± SD from three independent experiments. *P<0.05. FUBP1, far upstream element-binding protein 1; si, small interfering RNA; NC, negative control; OE, overexpression.
Fig 3: FUBP1 promotes pancreatic adenocarcinoma cell migration and invasion. Transwell assays were used to analyze the migratory and invasive abilities of (A) PaTu8988 cells transfected with si-NC or si-FUBP1 and (B) SW1990 cells transfected with FUBP1-OE or empty vectors (magnification, ×200). (C and E) The number of migrated cells was counted and analyzed. (D and F) Number of invasive cells was counted and analyzed. Wound healing assays were used to determine the cell migratory ability of (G) PaTu8988 cells transfected with si-NC or si-FUBP1 and (H) SW1990 cells transfected with empty or FUBP1-OE vectors (magnification, ×100). (I and J) The migratory distance was measured and analyzed. Data are presented as the mean ± SD from three independent experiments. *P<0.05. FUBP1, far upstream element-binding protein 1; si, small interfering RNA; NC, negative control; OE, overexpression.
Fig 4: FUBP1 promotes EMT in pancreatic adenocarcinoma cells. Western blotting was used to analyze the expression levels of the EMT-related proteins, E-cadherin, N-cadherin, ß-catenin and vimentin, in (A) PaTu8988 cells transfected with si-NC or si-FUBP1 and (B) SW1990 cells transfected with empty or FUBP1-OE vectors. ß-actin was used as the internal loading control. (C) Immunofluorescence assay was used to determine E-cadherin and vimentin expression levels in PaTu8988 cells transfected with si-NC or si-FUBP1 (magnification, ×200). Data are presented as the mean ± SD from three independent experiments. *P<0.05. FUBP1, far upstream element-binding protein 1; EMT, epithelial-mesenchymal transition; si, small interfering RNA; NC, negative control; OE, overexpression.
Fig 5: Upregulated FUBP1 expression levels are associated with a poor prognosis in patients with PAAD. (A) FUBP1 expression levels in PAAD and adjacent normal tissues from The Cancer Genome Atlas database. (B) starBase database was used to determine that, compared with patients with PAAD with low expression levels of FUBP1, patients with high expression levels of FUBP1 had a poorer overall survival (P=0.037). (C) mRNA expression levels of FUBP1 were upregulated in seven human PAAD tissues compared with adjacent normal tissues. Data are presented as the mean ± SD from three independent experiments. *P<0.05 and ***P<0.01. FUBP1, far upstream element-binding protein 1; PAAD, pancreatic adenocarcinoma.
Supplier Page from Abcam for Anti-FUBP1/FBP antibody [EPR12326]