mouse derived lamp1  (Proteintech)

Journal: Emerging microbes & infections

Article Title: GILT restricts the cellular entry mediated by the envelope glycoproteins of SARS-CoV, Ebola virus and Lassa fever virus.

doi: 10.1080/22221751.2019.1677446

Figure Lengend Snippet: Figure 1. GILT expression is upregulated by IFN-γ in lung cancer cells and has a lysosomal distribution. A549 and MRC-5 cells were treated with the indicated concentrations of IFN-α or IFN-γ for 12 h. (A) The levels of intracellular GILT and IFITM proteins were determined by Western blot assay. β-actin served as a loading control. (B) The localization of GILT in MRC-5 cells treated with IFN-γ was detected by immunofluorescent staining with anti-GILT polyclonal antibody (green). EEA1, Rab9 or LAMP1 were visual- ized by immunofluorescent staining with respective antibodies (red). Cell nuclei were stained blue with DAPI.

Article Snippet: Immunofluorescence To visualize the subcellular localization of wild-type and mutant GILT proteins, A549 or TRex 293 cells were first fixed with 4% paraformaldehyde for 20 min and then permeabilized with 0.1% Triton X-100 for 10 min. After incubation in the blocking solution (1×PBS with 3% BSA and 5% FBS) for 1 h, cells were stained with a mouse monoclonal antibody (G-11, SANTA CRUZ sc-393507A) or rabbit polyclonal antibody recognizing GILT protein (sc-393507A) together with a Rabbit derived monoclonal antibody against EEA1 (Cell Signaling, 2411), Rab9 (Cell Signaling, 5118s) or mouse derived LAMP1 monoclonal antibody (Cell Signaling, 15665), respectively.

Techniques: Expressing, Western Blot, Control, Staining

Journal: Emerging microbes & infections

Article Title: GILT restricts the cellular entry mediated by the envelope glycoproteins of SARS-CoV, Ebola virus and Lassa fever virus.

doi: 10.1080/22221751.2019.1677446

Figure Lengend Snippet: Figure 5. Mutations of N-linked glycosylation reduce GILT lysosomal localization. FLP-IN T Rex cells expressing the indicated wild- type and N-linked glycosylation mutant GILT proteins were treated with Tet for 24 h to induce GILT expression. The localization of wild-type GILT and its N-linked glycosylation mutants was detected by immunofluorescent staining with anti-GILT polyclonal anti- body (green). LAMP1 was visualized by immunofluorescent staining with anti-LAMP1 monoclonal antibody (red). Cell nuclei were stained blue with DAPI (blue). More than 10 fields were examined, and the representative images of one or a few cells are presented.

Article Snippet: Immunofluorescence To visualize the subcellular localization of wild-type and mutant GILT proteins, A549 or TRex 293 cells were first fixed with 4% paraformaldehyde for 20 min and then permeabilized with 0.1% Triton X-100 for 10 min. After incubation in the blocking solution (1×PBS with 3% BSA and 5% FBS) for 1 h, cells were stained with a mouse monoclonal antibody (G-11, SANTA CRUZ sc-393507A) or rabbit polyclonal antibody recognizing GILT protein (sc-393507A) together with a Rabbit derived monoclonal antibody against EEA1 (Cell Signaling, 2411), Rab9 (Cell Signaling, 5118s) or mouse derived LAMP1 monoclonal antibody (Cell Signaling, 15665), respectively.

Techniques: Glycoproteomics, Expressing, Mutagenesis, Staining

Journal: Emerging microbes & infections

Article Title: GILT restricts the cellular entry mediated by the envelope glycoproteins of SARS-CoV, Ebola virus and Lassa fever virus.

doi: 10.1080/22221751.2019.1677446

Figure Lengend Snippet: Figure 8. Schematic model for GILT-mediated restriction of viral entry in lysosome. Enveloped viruses enter cell at plasma mem- brane or escape from endocytic compartments via virus-host cell membrane fusion. While MLV that fuses at plasma membrane and vesicular stomatitis virus (VSV) and influenza A virus (IAV) that fuse at early endosome or late endosome are resistant to GILT- mediated inhibition, viruses including Ebola virus, Lassa virus and SARS-CoV, which fuse at lysosome, are restricted by GILT. NPC1: Niemann-Pick C1 protein (lysosomal receptor of Ebola virus); LAMP1: Lysosome-associated membrane glycoprotein 1 (lyso- somal receptor of Lassa virus).

Article Snippet: Immunofluorescence To visualize the subcellular localization of wild-type and mutant GILT proteins, A549 or TRex 293 cells were first fixed with 4% paraformaldehyde for 20 min and then permeabilized with 0.1% Triton X-100 for 10 min. After incubation in the blocking solution (1×PBS with 3% BSA and 5% FBS) for 1 h, cells were stained with a mouse monoclonal antibody (G-11, SANTA CRUZ sc-393507A) or rabbit polyclonal antibody recognizing GILT protein (sc-393507A) together with a Rabbit derived monoclonal antibody against EEA1 (Cell Signaling, 2411), Rab9 (Cell Signaling, 5118s) or mouse derived LAMP1 monoclonal antibody (Cell Signaling, 15665), respectively.

Techniques: Clinical Proteomics, Virus, Membrane, Inhibition