Journal: PLoS Pathogens

Article Title: Prion protein signaling induces M2 macrophage polarization and protects from lethal influenza infection in mice

doi: 10.1371/journal.ppat.1008823

Figure Lengend Snippet: ( A ) Western blotting (left panel) for total and phosphorylated SFK in the lungs from control IgG- and 38–2 mAb-treated mice at 3 dpi with 200 IFU of IAV/PR8. Actb, β-actin. The percent density of total SFK (middle panel) and SFK phosphorylated at tyrosine 416 (right panel) against the densities of the corresponding proteins in IgG-treated, infected lungs after being normalized by the densities of β-actin. **, p<0.01. ( B ) Survival rate (%, upper panels) and body weight loss (%, lower panels) of WT mice intraperitoneally administrated with control IgG (left panels) and 38–2 mAb (right panel) together with 20 mg/mouse of DS 1 day before intranasal infection with 200 IFU of IAV/PR8. Error bars, SD. ( C ) Western blotting (upper panel) for total SFK, phosphorylated SFK (Tyr416), the cleaved caspase 3, and the viral protein M2 in the lungs from control IgG/DMSO-, 38–2 mAb/DMSO-, control IgG/DS-, and 38–2 mAb/DS-treated mice at 3 dpi with 200 IFU of IAV/PR8. Actb, β-actin. Lungs from uninfected, saline/DMSO-treated mice were used as a control. The percent density (lower panels) of total SFK, phosphorylated SFK (Tyr416), the cleaved caspase 3, and the viral protein M2 against the densities of the corresponding proteins in uninfected, saline/DMSO-treated lungs after being normalized by the densities of β-actin. *, p<0.05; **, p<0.01.

Article Snippet: Anti-podoplanin antibody (Cat. No. D190-3) and anti-β-actin antibody (Cat. No. M177-3) from MBL (Nagoya, Japan;), anti-SP-C (Cat. No. sc-7706) and anti-CC10 antibodies (Cat. No. sc-9772) from Santa Cruz Biotechnology (Santa Cruz, CA), anti-pro-caspase 3 (Cat. No. 9662S), anti-cleaved caspase 3 (Cat. No. 9664S), anti-total SFK (Cat. No. 2109S) and anti-phosphorylated SFK (Tyr416) antibodies (Cat. No. 2101S) from Cell Signaling (Beverly, MA), anti-PB1 (Cat. No. GTX125923), anti-NS1 (Cat. No. GTX125990), anti-M2 (Cat. No. GTX125951) and anti-NP (Cat. No. GTX125989) antibodies from GeneTex (Irvine, CA), anti-SOD1 antibody (Cat. No. ab13498) from Abcam (Cambridge, UK), anti-CD3 (Cat. No. MAB4841), anti-MGL1/2 (Cat. No. AF4297) and HRP -conjugated anti-goat IgG antibodies (Cat. No. HAF017) from R&D systems (Minneapolis, MN), anti-MPO antibody from Proteintech (Rosemont, IL; Cat. No. 66177-1-Ig), PE anti-mouse/human CD11b antibody (Cat. No. 101207) from Biolegend (San Diego, CA), vitality full length hrGFP polyclonal antibody (Cat. No. 240141) from Agilent (Santa Clara, CA), HRP-conjugated anti-mouse IgG antibodies (Cat. No. NA931), HRP-conjugated anti-rabbit IgG antibodies (Cat. No. NA934), HRP-conjugated anti-rat IgG antibodies (Cat. No. NA935) from GE Healthcare (Buckinghamshire, UK), Alexa Fluor 596 donkey anti-mouse IgG antibody (Cat. No. A21203), Alexa Fluor 488 goat anti-rabbit IgG antibody (Cat. No. A11008), Alexa Fluor 596 donkey anti-goat IgG antibody (Cat. No. A11058), Texas Red-X goat anti-rat IgG antibody (Cat. No. T6392), and Alexa Fluor 647 goat anti-mouse antibody (Cat. No. A28181) from Invitrogen (Carlsbad, CA).

Techniques: Western Blot, Control, Infection, Saline

Journal: PLoS Pathogens

Article Title: Prion protein signaling induces M2 macrophage polarization and protects from lethal influenza infection in mice

doi: 10.1371/journal.ppat.1008823

Figure Lengend Snippet: ( A ) Double immunofluorescent staining for phosphorylated SFK (Tyr416) together with MGL1/2, CD3, and MPO in the lungs from control IgG- and 38–2 mAb-treated mice uninfected and at 3 dpi with 200 IFU of IAV/PR8. Un, uninfected. Bar, 200 μ m. ( B ) Fold mRNA expression (2 -ΔCt ) analyzed by real-time RT-PCR for IL-6, TNF-α, IFN-α, INF-γ, MCP-1, iNOS, IFIT1, MxA, ARG1, MGL1, and IL-10 in the lungs from control IgG/DMSO, control IgG/DS-, 38–2 mAb/DMSO-, and 38–2 mAb/DS-treated mice uninfected and at 3 dpi with 200 IFU of IAV/PR8 (n = 3 in each group). Un, uninfected.*, p<0.05;**, p<0.01.

Article Snippet: Anti-podoplanin antibody (Cat. No. D190-3) and anti-β-actin antibody (Cat. No. M177-3) from MBL (Nagoya, Japan;), anti-SP-C (Cat. No. sc-7706) and anti-CC10 antibodies (Cat. No. sc-9772) from Santa Cruz Biotechnology (Santa Cruz, CA), anti-pro-caspase 3 (Cat. No. 9662S), anti-cleaved caspase 3 (Cat. No. 9664S), anti-total SFK (Cat. No. 2109S) and anti-phosphorylated SFK (Tyr416) antibodies (Cat. No. 2101S) from Cell Signaling (Beverly, MA), anti-PB1 (Cat. No. GTX125923), anti-NS1 (Cat. No. GTX125990), anti-M2 (Cat. No. GTX125951) and anti-NP (Cat. No. GTX125989) antibodies from GeneTex (Irvine, CA), anti-SOD1 antibody (Cat. No. ab13498) from Abcam (Cambridge, UK), anti-CD3 (Cat. No. MAB4841), anti-MGL1/2 (Cat. No. AF4297) and HRP -conjugated anti-goat IgG antibodies (Cat. No. HAF017) from R&D systems (Minneapolis, MN), anti-MPO antibody from Proteintech (Rosemont, IL; Cat. No. 66177-1-Ig), PE anti-mouse/human CD11b antibody (Cat. No. 101207) from Biolegend (San Diego, CA), vitality full length hrGFP polyclonal antibody (Cat. No. 240141) from Agilent (Santa Clara, CA), HRP-conjugated anti-mouse IgG antibodies (Cat. No. NA931), HRP-conjugated anti-rabbit IgG antibodies (Cat. No. NA934), HRP-conjugated anti-rat IgG antibodies (Cat. No. NA935) from GE Healthcare (Buckinghamshire, UK), Alexa Fluor 596 donkey anti-mouse IgG antibody (Cat. No. A21203), Alexa Fluor 488 goat anti-rabbit IgG antibody (Cat. No. A11008), Alexa Fluor 596 donkey anti-goat IgG antibody (Cat. No. A11058), Texas Red-X goat anti-rat IgG antibody (Cat. No. T6392), and Alexa Fluor 647 goat anti-mouse antibody (Cat. No. A28181) from Invitrogen (Carlsbad, CA).

Techniques: Staining, Control, Expressing, Quantitative RT-PCR

Journal: PLoS Pathogens

Article Title: Prion protein signaling induces M2 macrophage polarization and protects from lethal influenza infection in mice

doi: 10.1371/journal.ppat.1008823

Figure Lengend Snippet: ( A ) Uncropped, full picture of Western blotting for PrP C with 38–2 mAb in the lungs (Lg) and peritoneal macrophages (PM) from WT and Prnp 0/0 mice. Actb, β-actin. ( B ) Western blotting for total SFK, phosphorylated SFK (Tyr416), and MGL1/2 in peritoneal macrophages from WT and Prnp 0/0 mice 3 hrs after treatment with control IgG and 38–2 mAb together with DS or control DMSO. ( C ) ELISA for INF-γ, TNF-α, IL-4, and IL-10 in the culture medium from WT peritoneal macrophages at 0.5, 4, 16, and 24 hours (hrs) after treatment with control IgG and 38–2 mAb.*, p<0.05;** p<0.01. ( D ) Western blotting for total SFK, phosphorylated SFK (Tyr416), and MGL1/2 in peritoneal macrophages (PM) from WT and Prnp 0/0 mice 1 and 3 days after peritoneal injection with control IgG and 38–2 mAb together with control DMSO and DS.

Article Snippet: Anti-podoplanin antibody (Cat. No. D190-3) and anti-β-actin antibody (Cat. No. M177-3) from MBL (Nagoya, Japan;), anti-SP-C (Cat. No. sc-7706) and anti-CC10 antibodies (Cat. No. sc-9772) from Santa Cruz Biotechnology (Santa Cruz, CA), anti-pro-caspase 3 (Cat. No. 9662S), anti-cleaved caspase 3 (Cat. No. 9664S), anti-total SFK (Cat. No. 2109S) and anti-phosphorylated SFK (Tyr416) antibodies (Cat. No. 2101S) from Cell Signaling (Beverly, MA), anti-PB1 (Cat. No. GTX125923), anti-NS1 (Cat. No. GTX125990), anti-M2 (Cat. No. GTX125951) and anti-NP (Cat. No. GTX125989) antibodies from GeneTex (Irvine, CA), anti-SOD1 antibody (Cat. No. ab13498) from Abcam (Cambridge, UK), anti-CD3 (Cat. No. MAB4841), anti-MGL1/2 (Cat. No. AF4297) and HRP -conjugated anti-goat IgG antibodies (Cat. No. HAF017) from R&D systems (Minneapolis, MN), anti-MPO antibody from Proteintech (Rosemont, IL; Cat. No. 66177-1-Ig), PE anti-mouse/human CD11b antibody (Cat. No. 101207) from Biolegend (San Diego, CA), vitality full length hrGFP polyclonal antibody (Cat. No. 240141) from Agilent (Santa Clara, CA), HRP-conjugated anti-mouse IgG antibodies (Cat. No. NA931), HRP-conjugated anti-rabbit IgG antibodies (Cat. No. NA934), HRP-conjugated anti-rat IgG antibodies (Cat. No. NA935) from GE Healthcare (Buckinghamshire, UK), Alexa Fluor 596 donkey anti-mouse IgG antibody (Cat. No. A21203), Alexa Fluor 488 goat anti-rabbit IgG antibody (Cat. No. A11008), Alexa Fluor 596 donkey anti-goat IgG antibody (Cat. No. A11058), Texas Red-X goat anti-rat IgG antibody (Cat. No. T6392), and Alexa Fluor 647 goat anti-mouse antibody (Cat. No. A28181) from Invitrogen (Carlsbad, CA).

Techniques: Western Blot, Control, Enzyme-linked Immunosorbent Assay, Injection

Journal: PLoS Pathogens

Article Title: Prion protein signaling induces M2 macrophage polarization and protects from lethal influenza infection in mice

doi: 10.1371/journal.ppat.1008823

Figure Lengend Snippet: ( A ) Western blotting for total SFK, phosphorylated SFK (Tyr416), and MGL1/2 in peritoneal macrophages from WT and Prnp 0/0 mice 3 hrs after treatment with control IgG and 3S9 and 2H9 mAbs together with control DMSO and DS. ( B ) Survival rate (%, upper panels) and body weight loss (%, lower panels) of WT mice intraperitoneally administered with control IgG and 3S9 and 2H9 mAbs 1 day before intranasal infection with 100 (left panels) or 200 IFU (right panels) of IAV/PR8. Error bars, SD. *, p<0.05; **, p<0.01; IgG vs 3S9. #, <0.05; IgG vs 2H9.

Article Snippet: Anti-podoplanin antibody (Cat. No. D190-3) and anti-β-actin antibody (Cat. No. M177-3) from MBL (Nagoya, Japan;), anti-SP-C (Cat. No. sc-7706) and anti-CC10 antibodies (Cat. No. sc-9772) from Santa Cruz Biotechnology (Santa Cruz, CA), anti-pro-caspase 3 (Cat. No. 9662S), anti-cleaved caspase 3 (Cat. No. 9664S), anti-total SFK (Cat. No. 2109S) and anti-phosphorylated SFK (Tyr416) antibodies (Cat. No. 2101S) from Cell Signaling (Beverly, MA), anti-PB1 (Cat. No. GTX125923), anti-NS1 (Cat. No. GTX125990), anti-M2 (Cat. No. GTX125951) and anti-NP (Cat. No. GTX125989) antibodies from GeneTex (Irvine, CA), anti-SOD1 antibody (Cat. No. ab13498) from Abcam (Cambridge, UK), anti-CD3 (Cat. No. MAB4841), anti-MGL1/2 (Cat. No. AF4297) and HRP -conjugated anti-goat IgG antibodies (Cat. No. HAF017) from R&D systems (Minneapolis, MN), anti-MPO antibody from Proteintech (Rosemont, IL; Cat. No. 66177-1-Ig), PE anti-mouse/human CD11b antibody (Cat. No. 101207) from Biolegend (San Diego, CA), vitality full length hrGFP polyclonal antibody (Cat. No. 240141) from Agilent (Santa Clara, CA), HRP-conjugated anti-mouse IgG antibodies (Cat. No. NA931), HRP-conjugated anti-rabbit IgG antibodies (Cat. No. NA934), HRP-conjugated anti-rat IgG antibodies (Cat. No. NA935) from GE Healthcare (Buckinghamshire, UK), Alexa Fluor 596 donkey anti-mouse IgG antibody (Cat. No. A21203), Alexa Fluor 488 goat anti-rabbit IgG antibody (Cat. No. A11008), Alexa Fluor 596 donkey anti-goat IgG antibody (Cat. No. A11058), Texas Red-X goat anti-rat IgG antibody (Cat. No. T6392), and Alexa Fluor 647 goat anti-mouse antibody (Cat. No. A28181) from Invitrogen (Carlsbad, CA).

Techniques: Western Blot, Control, Infection