Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-0

Ubated with virus-containing medium (MOI = 5) at 37°C for 1 h, medium was replaced, and cells were further incubated at 37°C for the times as indicated. (c) A549 cells were incubated with virus-containing medium (MOI = 5) at 4°C for 20 min and then at 37°C for the times as indicated. CK2β activation was assessed at the indicated time points by Western blot with polyclonal rabbit anti-phosphorylated CK2β (P-CK2β). Bands in three independent experiments were quantified, and relative CK2β activation (black bars) was calculated and normalized to the loading control (anti-CK2β mAb). White bars represent uninoculated cells.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-7

Ubated with virus-containing medium (MOI = 5) at 37°C for 1 h, medium was replaced, and cells were further incubated at 37°C for the times as indicated. (c) A549 cells were incubated with virus-containing medium (MOI = 5) at 4°C for 20 min and then at 37°C for the times as indicated. CK2β activation was assessed at the indicated time points by Western blot with polyclonal rabbit anti-phosphorylated CK2β (P-CK2β). Bands in three independent experiments were quantified, and relative CK2β activation (black bars) was calculated and normalized to the loading control (anti-CK2β mAb). White bars represent uninoculated cells.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-1

.t.) by Western blot analysis with a mAb specific for the β-subunit of CK2. Loading was controlled with a mAb against β-actin. (b) Total CK2α protein detected 24 h, 48 h, and 72 h after transfection with CK2β siRNA.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-6

Rol or CK2β siRNA for 48 h, then co-transfected with plasmids containing the PB2, PB1, PA, and NP genes plus a luciferase reporter plasmid. Cells not transfected with the PB1 plasmid were used as negative controls. After 24 h, polymerase (luciferase) activity was assayed in cell extracts. Results represent the mean ± SE of 3 independent experiments.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-3

Ubated at 37°C for 30 min. Influenza A virions (red) were detected with a goat anti-IVA (H1N1) polyclonal antibody, and an epidermal growth factor receptor (EGFR)-specific mAb (green) was used as a cell membrane marker. Images were taken with a confocal laser scanning microscope at 100× magnification. Blue arrows indicate virus accumulation at the nuclear membrane; yellow arrows indicate virus accumulation in the cytoplasm and at the cell membrane.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-5

h. The RNP complexes and CK2β protein were stained with a goat anti-NP antibody (red) and an anti-CK2β mouse monoclonal antibody (green), respectively. The nucleus was counterstained with TO-PRO-3 (blue). Intracellular RNP localization was analyzed at the indicated time points by confocal laser scanning microscopy (100× magnification).<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p

Beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content-4

Bated for 9 h or 24 h before Western blot analysis. Viral proteins were detected with a goat anti-IVA (H1N1) polyclonal antibody. Loading was controlled with a mAb against β-actin. Bands of viral protein were quantified as a percentage of control values normalized to the loading control. Shown are the mean ± SE from 3 independent experiments. (b) Cells were treated as above. The percentage of NP-expressing cells was measured at 24 h p.i. by flow cytometry (FACS) using an anti-NP mAb. The experiments were performed in duplicate.<p><b>Copyright information:</b></p><p>Taken from "beta gene silencing increases cell susceptibility to influenza A virus infection resulting in accelerated virus entry and higher viral protein content"</p><p>http://www.jmolecularsignaling.com/content/3/1/13</p><p>Journal of Molecular Signaling 2008;3():13-13.</p><p>Published online 23 Jul 2008</p><p>PMCID:PMC2494991.</p><p></p