Fig 1: The involvement of FIP2 in human macrophage biology.(A) Gene Ontology mapping showing the effect of FIP2 silencing on biological processes in primary human macrophages from 7 donors. (B) Effect of FIP2 silencing on E. coli-stimulated induction of IRF3-target genes. (C) Effect of FIP2 silencing on the E. coli-stimulated induction of a selection of pro-inflammatory genes. Data are median counts with boxed 1.5 IQR 95%. **** (FDR< 0.0001), *** (FDR<0.001), ** (FDR<0.01), * (FDR<0.05). FDR = False Discovery Rate, IQR = Inter Quantile Range.
Fig 2: Activation of RIG-I signal pathway is inhibited by miR-9-mimic transfection. RD cells were transfected with miR-9-mimic and then infected with EV71 at 24 h posttransfection. (A) Expression of RIG-I, NF?B, phosphorylated NF?B, IRF3, and phosphorylated IRF3 of infected cells was examined by western blotting at several time points postinfection. (B) We performed real-time PCR to examine gene expression of NF?B, RIG-I, and IRF3 at several time points following infection. These assays have been performed three times. Data represent means ± SD. *P < 0.05; ***P < 0.001. The statistical test performed using Student's t-test.
Fig 3: FIP2 controls E. coli induced IFN-ß mRNA induction and secretion.(A) Immunoblots showing the phosphorylation patterns of TBK1, IRF3, I?Ba and p38MAPK in FIP2 silenced THP-1 cells stimulated with E. coli bioparticles or LPS (100 ng/ml). Data are representative of three independent experiments. (B) Quantification of phosphorylation patterns of the proteins shown in the immunoblots presented in (A). (C) ELISA quantification IFN-ß and TNF secretion in THP-1 cells treated with NS RNA or FIP2 siRNA and stimulated as indicated. (D) Quantification of E. coli-stimulated IFN-ß and TNF mRNAs in THP-1 cells with lentiviral overexpression of FIP2. (E) Quantification of Poly I:C and LPS stimulated IFN-ß mRNA induction in cells treated with NS RNA or FIP2 siRNA after 4 hours of stimulation. Poly I:C (5 µg/ml) was transfected using Lipofectamine® 2000. Data are representative of three independent experiments.
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