Journal: Virology Journal

Article Title: Classical swine fever virus triggers RIG-I and MDA5-dependent signaling pathway to IRF-3 and NF-κB activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages

doi: 10.1186/1743-422X-10-286

Figure Lengend Snippet: Expression and nuclear translocation of IRF-3 after CSFV infection in porcine alveolar macrophages. (A) Expression of IRF-3 was measured by Western Blotting with antibodies specific for IRF-3, and the cells were treated as demonstrated in Figure . (B) Indirect immunofluorescence analysis was used to measure cellular localization of IRF-3 in CSFV-infected PAMs. Cells were mock treated, poly (I:C) stimulated, or infected with Shimen isolates at an MOI of 3. At 24 hpi, cells were fixed and the localization of IRF-3 (red) was observed by fluorescence microscope using immunofluorescence stain with anti-IRF-3 and QDs-SA 605-conjugated and biotinylated secondary antibodies. Nuclei were stained with DAPI. Bar, 10 μm. The experiment was repeated three times and the figure shows a representative experiment. An asterisk indicates a statistically significant difference from uninfected cells, * P < 0.05 and ** P < 0.01.

Article Snippet: After washed three times with PBST, cells were further incubated with QDs-SA-conjugated and biotinylated secondary antibodies (1:100, Molecular Probes, Jiayuan Quantum Dots, Wuhai, China) at 37°C for 30 min.

Techniques: Expressing, Translocation Assay, Infection, Western Blot, Immunofluorescence, Fluorescence, Microscopy, Staining

Journal: Virology Journal

Article Title: Classical swine fever virus triggers RIG-I and MDA5-dependent signaling pathway to IRF-3 and NF-κB activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages

doi: 10.1186/1743-422X-10-286

Figure Lengend Snippet: Protein expression and nuclear translocation of NF- κB after CSFV infection in porcine alveolar macrophages. (A) Expression of NF-κB was measured by Western Blotting with antibodies specific for NF-κB, the cells were treated as demonstrated in Figure . (B) Indirect immunofluorescence analysis was used to measure cellular localization of NF-κB in CSFV-infected PAMs. Cells were treated as indicated in Figure . At 24 hpi, cells were fixed and the localization of NF-κB (red) was observed by fluorescence microscope using immunofluorescence stain with anti-NF-κB/p65 and QDs-SA 605-conjugated biotinylated secondary antibodies. Nuclei were stained with DAPI. Bar, 10 μm. Representative results are shown of one of three separate experiments. An asterisk indicates a statistically significant difference from uninfected cells, * P < 0.05 and ** P < 0.01.

Article Snippet: After washed three times with PBST, cells were further incubated with QDs-SA-conjugated and biotinylated secondary antibodies (1:100, Molecular Probes, Jiayuan Quantum Dots, Wuhai, China) at 37°C for 30 min.

Techniques: Expressing, Translocation Assay, Infection, Western Blot, Immunofluorescence, Fluorescence, Microscopy, Staining