Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Influenza Virus Infection Induces ZBP1 Expression and Necroptosis in Mouse Lungs

doi: 10.3389/fcimb.2019.00286

Figure Lengend Snippet: Immunohistochemical detection of ZBP1 in IAV-infected mouse lungs. Lung sections from saline-treated (Mock, A , B ) or H1N1 PR8-infected mice on day 7 post-infection (C,D) were subjected to immunostaining with a ZBP1 monoclonal antibody by using Vector M.O.M immunodetection kits ( n = 5). Positive immunoreactivities (red) were detected in the alveolar epithelial cells (red arrows) and immune cells (green arrows). The region indicated in (C) (magnification, x100) is shown at higher magnification in (D) (x400).

Article Snippet: Vector MOM immunodetection kit (PK-2200), Vector NovaRED substrate (SK-4800), Vector hematoxylin (H-3401), VectaMount permanent mounting medium (H-5000), and Vectashield hardset antifade mounting medium with DAPI (H-1500) were from Vector Laboratories (Burlingame, CA).

Techniques: Immunohistochemical staining, Infection, Immunostaining, Plasmid Preparation, Immunodetection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Influenza Virus Infection Induces ZBP1 Expression and Necroptosis in Mouse Lungs

doi: 10.3389/fcimb.2019.00286

Figure Lengend Snippet: IAV induces phosphorylation of MLKL in alveolar epithelial cells and infiltrated immune cells in infected mouse lungs. Lung sections from saline-treated (Mock, A , B ) or PR8-infected mice on day 7 post-infection (C–F) were subjected to immunostaining with a phospho-MLKL (Ser345) monoclonal antibody using a Vector M.O.M immunodetection kit ( n = 6). Positive p-MLKL immunoreactivities (red) were detected in the alveolar epithelial cells (red arrows) and immune cells (green arrows). The region indicated in (E) (magnification, x100) is shown at higher magnification in (F) (x400). Final magnification for (A) is x100 and for (B–D) x400. In (A–D) , the lung sections were from inflated mouse lungs by intratracheal infusion of 10% formalin to total lung capacity.

Article Snippet: Vector MOM immunodetection kit (PK-2200), Vector NovaRED substrate (SK-4800), Vector hematoxylin (H-3401), VectaMount permanent mounting medium (H-5000), and Vectashield hardset antifade mounting medium with DAPI (H-1500) were from Vector Laboratories (Burlingame, CA).

Techniques: Infection, Immunostaining, Plasmid Preparation, Immunodetection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Influenza Virus Infection Induces ZBP1 Expression and Necroptosis in Mouse Lungs

doi: 10.3389/fcimb.2019.00286

Figure Lengend Snippet: Immunofluorescence detection of IAV-induced phosphorylation of MLKL and its co-localization with pulmonary infiltrated immune cells. (A,B) Lung sections from control (A) or PR8-infected mice (B) on day 7 post-infection were subjected to immunofluorescence staining with a phospho-MLKL (Ser345) monoclonal antibody by using Vector M.O.M immunodetection kit followed by Alexa Fluor 568-labeled goat anti-mouse secondary IgG ( n = 6). Positive p-MLKL immunoreactivities are stained red (indicated by green arrows) and cell nuclei stained blue by DAPI. Final magnification is x400. (C,D) Lung sections from PR8-infected mice on day 7 post-infection were subjected to double immunofluorescence staining with the phospho-MLKL (Ser345) mouse monoclonal antibody, Alexa Fluor-488 conjugated CD45 or FITC conjugated CD192/CCR2 rat monoclonal antibodies as described in section Materials and Methods. Positive p-MLKL immunoreactivities are stained red, and CD45 or CCR2 are stained green. The co-localization or overlay is shown as yellow and indicated by white arrows. Final magnification: x400 ( n = 6).

Article Snippet: Vector MOM immunodetection kit (PK-2200), Vector NovaRED substrate (SK-4800), Vector hematoxylin (H-3401), VectaMount permanent mounting medium (H-5000), and Vectashield hardset antifade mounting medium with DAPI (H-1500) were from Vector Laboratories (Burlingame, CA).

Techniques: Immunofluorescence, Infection, Staining, Plasmid Preparation, Immunodetection, Labeling, Double Immunofluorescence Staining