Journal: PLOS One

Article Title: Down-regulation of circ0001361 induces apoptosis and suppresses the progression of glioma

doi: 10.1371/journal.pone.0343681

Figure Lengend Snippet: (A) MiRNAs targeted by circ0001361 were predicted. (B) Expression of hsa-miR-525-5p in glioma (Group A) and normal brain tissue (Group B) was shown. (C) Expression of hsa-miR-525-5p in glioma grade Ⅱ -Ⅳ was shown. (D) Prognostic significance of hsa-miR-525-5p was analyzed. (E) Potential binding sites between circ0001361 and hsa-miR-525-5p were predicted. (F) Circ0001361-hsa-miR-525-5p interaction was confirmed by dual-luciferase reporter assay.

Article Snippet: After 48 hours, the Firefly luciferase and Renilla Luciferase were detected using the Dual Luciferase Reporter Assay Kit (Vazyme, China).

Techniques: Expressing, Binding Assay, Luciferase, Reporter Assay

Journal: PLOS One

Article Title: Down-regulation of circ0001361 induces apoptosis and suppresses the progression of glioma

doi: 10.1371/journal.pone.0343681

Figure Lengend Snippet: (A) Genes targeted by hsa-miR-525-5p were predicted. (B) Correlation between hsa-miR-525-5p and MEIS1 was analyzed. (C) Prognostic significance of MEIS1 was analyzed. (D) Expression of MEIS1 in GBM, LGG and normal brain tissues was shown. (E) Expression of MEIS1 in glioma grade Ⅱ -Ⅳ was shown. (F) The relationship between MEIS1 and IDH mutation status was shown. (G) The relationship between MEIS1 and 1p/19q co−deletion status was shown. (H) Potential binding sites between hsa-miR-525-5p and MEIS1 were predicted. (I) Hsa-miR-525-5p and MEIS1 interaction was confirmed by dual-luciferase reporter assay.

Article Snippet: After 48 hours, the Firefly luciferase and Renilla Luciferase were detected using the Dual Luciferase Reporter Assay Kit (Vazyme, China).

Techniques: Expressing, Mutagenesis, Binding Assay, Luciferase, Reporter Assay

Journal: bioRxiv

Article Title: Epoxyeicosatrienoic acids and sEH inhibition prevent cardiac dysfunction in CVB3-induced myocarditis by positively regulating type I interferon signaling

doi: 10.1101/2023.02.03.527086

Figure Lengend Snippet: ( A ) Phosphorylation of IRF3 (on Ser396) in uninfected (CON) or CVB3-infected AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=4). ( B ) Dimerization of IRF3 in uninfected (CON) or CVB3-infected AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=4). ( C ) Nuclear translocation of IRF3 in uninfected (CON) or CVB3-infected AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=4). GAPDH and LaminA/C were used as cytoplasm and nuclear protein loading controls. ( D ) Immunofluorescence images showing the impact of 14,15-EET and TPPU on the subcellular localization of IRF3 in uninfected (CON) or CVB3-infected AC16 cells; bars =50 μm. ( E ) Scheme showing the two main RNA sensor-induced IRF3 activation pathways. ( F-G ) AC16 cells that were uninfected (CON) or infected with CVB3 were treated with 11,12-EET, 14,15-EET or TPPU (n=3). Shown are changes in IRF3 phosphorylation following the siRNA-mediated down regulation of MDA5 (F), or TLR3 (G). ( H ) Activity of a IFNβ-luciferase reporter construct in HEK293 cells expressing either a control vector (vector), pcMDA5, pcMAVS, pcTBK1 , or pcIRF3-5D and treated with solvent (CON) or 14,15-EET (1 μM) for 24 hours. ( I ) Impact of the siRNA-mediated downregulation of TBK1 on the phosphorylation of IRF3 (on Ser396) in uninfected (CON) or CVB3-infected AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=3). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: After 48 hours, transfected HEK293 cells were harvested in immunoprecipitation (IP) buffer containing 20 mM Tris (pH7.5), 150 mM NaCl, 1% Triton X-100 with freshly added PMSF, protease inhibitors and phosphatase inhibitors (Boster Biological Tech, Wuhan, China).

Techniques: Phospho-proteomics, Infection, Translocation Assay, Immunofluorescence, Activation Assay, Activity Assay, Luciferase, Construct, Expressing, Control, Plasmid Preparation, Solvent

Journal: bioRxiv

Article Title: Epoxyeicosatrienoic acids and sEH inhibition prevent cardiac dysfunction in CVB3-induced myocarditis by positively regulating type I interferon signaling

doi: 10.1101/2023.02.03.527086

Figure Lengend Snippet: ( A ) Impact of the siRNA-mediated downregulationof GSK3β on the phosphorylation of IRF3 (on Ser396) in uninfected (CON) or CVB3-infected AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=4). ( B ) Impact of the EET antagonist; EEZE (1 μM) on the phosphorylation of GSK3β in AC16 cells treated with 11,12-EET, 14,15-EET or TPPU (n=4). ( C ) Co-immunoprecipitation of TBK1 with a GSK3β-Flag fusion protein from HEK293 cells treated with 14,15-EET (1 μM) or TPPU (10 μM) in the absence or presence of EEZE (n=3). ( D ) Impact of the wild-type GSK3β and the Y216F and Y216D GSK3β mutants on the phosphorylation of TBK1 in uninfected and CVB3-infected HEK293 cells (n=3). ( E-F ) Impact of GSK3β mutation alone and in combination with 14,15-EET on the phosphorylation of TBK1 in uninfected and CVB3-infected HEK293 cells (n=3-4). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: After 48 hours, transfected HEK293 cells were harvested in immunoprecipitation (IP) buffer containing 20 mM Tris (pH7.5), 150 mM NaCl, 1% Triton X-100 with freshly added PMSF, protease inhibitors and phosphatase inhibitors (Boster Biological Tech, Wuhan, China).

Techniques: Phospho-proteomics, Infection, Immunoprecipitation, Mutagenesis