Journal: PLoS ONE

Article Title: Mumps Virus Induces Protein-Kinase-R-Dependent Stress Granules, Partly Suppressing Type III Interferon Production

doi: 10.1371/journal.pone.0161793

Figure Lengend Snippet: FL cells were transfected with targeting siRNA (G3BP1 si and TIA-1 si) or nontargeting control siRNA (N. C. si) for 48 h. (A) Immunofluorescence staining of IGF2BP1 (green) and MuV F (red) proteins and staining with DAPI (blue). Scale bar indicates 10 μm. (B) Quantification of foci containing IGF2BP1 in MuV-infected cells observed by immunofluorescence microscopy ( n = 3). F-protein-positive cells were counted as MuV-infected cells. (C) Real-time RT-PCR analysis in G3BP1 and TIA-1 double-KD cells and control cells transfected with G3BP1 siRNA/TIA-1 siRNA and N.C. siRNA, respectively, at 48 h post-transfection ( n = 3). (D) Phosphorylation status of PKR, eIF2α and IRF3 analyzed by western blotting. β-Actin was determined as a control. (E) Amounts of IFN-β and IFN-λ1 in culture supernatants ( n = 4) at 24 h post-infection determined by ELISA. (F) Viral titer in supernatants ( n = 4) at 24 h post-infection. The bar graphs represent means ± standard deviation. * P <0.05, ** P <0.01, N.S.: not significant.

Article Snippet: Rabbit PcAbs against IRF3 (309033), RIG-I (PAB12973), melanoma differentiation-associated gene 5 (MDA5) (29020), Hu-antigen R (HuR), and insulin-like growth factor 2 binding protein (IGF2BP)1 were purchased from Active Motif (Carlsbad, CA), Abnova (Taipei, Taiwan), Immuno-Biological Laboratories (Gunma, Japan), Medical and Biological Laboratory (Nagoya, Japan) and Abgent (San Diego, CA), respectively.

Techniques: Transfection, Control, Immunofluorescence, Staining, Infection, Microscopy, Quantitative RT-PCR, Phospho-proteomics, Western Blot, Enzyme-linked Immunosorbent Assay, Standard Deviation

Journal: PLoS ONE

Article Title: Mumps Virus Induces Protein-Kinase-R-Dependent Stress Granules, Partly Suppressing Type III Interferon Production

doi: 10.1371/journal.pone.0161793

Figure Lengend Snippet: BEAS-2 cells were transfected with targeting siRNA (PKR si, or G3BP1 si/TAI-1 si), or nontargeting control siRNA (N.C. si) for 48 h. Cells were infected with RSV at MOI 10. (A) Immunostaining with RSV proteins (green) and G3BP1 (red), and staining with DAPI (blue) of the cells at 24 h post-infection. Scale bar indicates 5 μm. (B) Amounts of IFN-β and IFN-λ1 in the culture supernatants ( n = 4) at 24 h post-infection, determined by ELISA. (C) Expression of RSV proteins, and phosphorylation status of PKR, eIF2α and IRF3 determined by western blotting. β-Actin was examined as a control. The bar graphs represent means ± standard deviation. * P <0.05, ** P <0.01 versus RSV-infected control (N.C. siRNA transfected) cells.

Article Snippet: Rabbit PcAbs against IRF3 (309033), RIG-I (PAB12973), melanoma differentiation-associated gene 5 (MDA5) (29020), Hu-antigen R (HuR), and insulin-like growth factor 2 binding protein (IGF2BP)1 were purchased from Active Motif (Carlsbad, CA), Abnova (Taipei, Taiwan), Immuno-Biological Laboratories (Gunma, Japan), Medical and Biological Laboratory (Nagoya, Japan) and Abgent (San Diego, CA), respectively.

Techniques: Transfection, Control, Infection, Immunostaining, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Phospho-proteomics, Western Blot, Standard Deviation