hiv 1 infection  (Sino Biological)


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    Name:
    Human Immunodeficiency Virus type 1 gp120 Glycoprotein 120 ELISA Kit
    Description:
    The kit has been verified by high purity Human Immunodeficiency Virus type 1 HIV 1 gp120 Glycoprotein 120 recombinant protein
    Catalog Number:
    KIT11233
    Price:
    None
    Category:
    ELISA Kit
    Reactivity:
    HIV
    Buy from Supplier


    Structured Review

    Sino Biological hiv 1 infection
    NSC- and A549-derived exosomes significantly enhance <t>HIV-1</t> entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P
    The kit has been verified by high purity Human Immunodeficiency Virus type 1 HIV 1 gp120 Glycoprotein 120 recombinant protein
    https://www.bioz.com/result/hiv 1 infection/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hiv 1 infection - by Bioz Stars, 2021-06
    93/100 stars

    Images

    1) Product Images from "Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells"

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells

    Journal: International Journal of Nanomedicine

    doi: 10.2147/IJN.S132762

    NSC- and A549-derived exosomes significantly enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P
    Figure Legend Snippet: NSC- and A549-derived exosomes significantly enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P

    Techniques Used: Derivative Assay, Expressing, Luciferase, Activity Assay, Infection

    Breast milk- and plasma-derived exosomes enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) breast milk-derived exosomes (0.035 μg) or ( C ) plasma-derived exosomes (0.05 μg). Virus entry was evaluated in the presence of exosomes and anti-TIM-4 antibodies. ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) breast milk-derived exosomes (0.035 μg) or ( D ) plasma-derived exosomes (0.05 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA. ** P
    Figure Legend Snippet: Breast milk- and plasma-derived exosomes enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) breast milk-derived exosomes (0.035 μg) or ( C ) plasma-derived exosomes (0.05 μg). Virus entry was evaluated in the presence of exosomes and anti-TIM-4 antibodies. ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) breast milk-derived exosomes (0.035 μg) or ( D ) plasma-derived exosomes (0.05 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA. ** P

    Techniques Used: Derivative Assay, Expressing, Luciferase, Activity Assay, Infection

    Related Articles

    Cell Culture:

    Article Title: The Neuropeptides Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Control HIV-1 Infection in Macrophages Through Activation of Protein Kinases A and C
    Article Snippet: .. HIV-1 replication was quantified in cell culture supernatants after 10–12 days of infection by a commercial ELISA kit (Sino Biological), according to manufacturer’s instructions. .. HIV-1-infected macrophages were treated either with VIP or PACAP immediately after cell infection, or, in some assays, before infection, and maintained during culture.

    Infection:

    Article Title: The Neuropeptides Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Control HIV-1 Infection in Macrophages Through Activation of Protein Kinases A and C
    Article Snippet: .. HIV-1 replication was quantified in cell culture supernatants after 10–12 days of infection by a commercial ELISA kit (Sino Biological), according to manufacturer’s instructions. .. HIV-1-infected macrophages were treated either with VIP or PACAP immediately after cell infection, or, in some assays, before infection, and maintained during culture.

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells
    Article Snippet: .. Blocking of HIV-1 infection A protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control. .. Blocking of HIV-1 infectionA protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control.

    Enzyme-linked Immunosorbent Assay:

    Article Title: The Neuropeptides Vasoactive Intestinal Peptide and Pituitary Adenylate Cyclase-Activating Polypeptide Control HIV-1 Infection in Macrophages Through Activation of Protein Kinases A and C
    Article Snippet: .. HIV-1 replication was quantified in cell culture supernatants after 10–12 days of infection by a commercial ELISA kit (Sino Biological), according to manufacturer’s instructions. .. HIV-1-infected macrophages were treated either with VIP or PACAP immediately after cell infection, or, in some assays, before infection, and maintained during culture.

    Derivative Assay:

    Article Title: Comparison of the Specificities of IgG, IgG-Subclass, IgA and IgM Reactivities in African and European HIV-Infected Individuals with an HIV-1 Clade C Proteome-Based Array
    Article Snippet: .. HIV-1 clade C derived recombinant gp120 (rgp120) expressed in 293 cells, was purchased from Sino Biological (Beijing, People’s Republic of China) [ ]. .. The identity of the proteins was verified by SDS-PAGE and Coomassie staining, as well as by Westernblot (i.e. with mouse a-His-tag antibody, Dianova, Hamburg, Germany, and alkaline-phosphatase-coupled rabbit anti-mIgG, BD, Franklin Lakes, NJ, USA) and by mass spectrometry (Microflex MALDI-TOF, Bruker).

    Recombinant:

    Article Title: Comparison of the Specificities of IgG, IgG-Subclass, IgA and IgM Reactivities in African and European HIV-Infected Individuals with an HIV-1 Clade C Proteome-Based Array
    Article Snippet: .. HIV-1 clade C derived recombinant gp120 (rgp120) expressed in 293 cells, was purchased from Sino Biological (Beijing, People’s Republic of China) [ ]. .. The identity of the proteins was verified by SDS-PAGE and Coomassie staining, as well as by Westernblot (i.e. with mouse a-His-tag antibody, Dianova, Hamburg, Germany, and alkaline-phosphatase-coupled rabbit anti-mIgG, BD, Franklin Lakes, NJ, USA) and by mass spectrometry (Microflex MALDI-TOF, Bruker).

    Article Title: Tunable Fano-Resonant Metasurfaces on a Disposable Plastic-Template for Multimodal and Multiplex Biosensing.
    Article Snippet: .. Metasurfaces are engineered nanostructured interfaces that extend the photonic behavior of natural materials, and they spur many breakthroughs in multiple fields, including quantum optics, optoelectronics, and biosensing. .. Metasurfaces are engineered nanostructured interfaces that extend the photonic behavior of natural materials, and they spur many breakthroughs in multiple fields, including quantum optics, optoelectronics, and biosensing.

    Blocking Assay:

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells
    Article Snippet: .. Blocking of HIV-1 infection A protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control. .. Blocking of HIV-1 infectionA protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control.

    Incubation:

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells
    Article Snippet: .. Blocking of HIV-1 infection A protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control. .. Blocking of HIV-1 infectionA protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control.

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    Sino Biological hiv 1 infection
    NSC- and A549-derived exosomes significantly enhance <t>HIV-1</t> entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P
    Hiv 1 Infection, supplied by Sino Biological, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv 1 infection/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hiv 1 infection - by Bioz Stars, 2021-06
    93/100 stars
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    94
    Sino Biological rabbit anti gp120 polyclonal antibody
    Effects of GPI-anchored antibodies on HIV-1 Env processing. A The expression levels of gp160, <t>gp120,</t> gp41, P24, and β-actin in HEK293T cells coexpressing GPI-m36.4 or GPI-FluIgG03 and NL4-3 (left panel) or THRO.c/2626 (right panel) were detected by Western blotting. For clarity, images were spliced and grouped, and the original images are provided in the Supplementary materials . B The ratio of gp120 to gp160 in cell lysates was determined by quantifying the corresponding band intensities with ImageJ. The data shown were derived from three independent experiments, and error bars indicate standard deviations. Statistical comparisons were conducted by ANOVA (** P
    Rabbit Anti Gp120 Polyclonal Antibody, supplied by Sino Biological, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rabbit anti gp120 polyclonal antibody/product/Sino Biological
    Average 94 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    rabbit anti gp120 polyclonal antibody - by Bioz Stars, 2021-06
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    95
    Sino Biological commercial p24 elisa kit
    PSGL-1 inhibits Env incorporation in virions and viral entry. a Virions harvested from producer 293 T cells transfected with two different doses of PSGL-1 or an empty vector were pelleted through 20% sucrose cushion were analyzed by Western blotting together with the cell lysates. b PSGL-1-knockout Jurkat cells lines with two different sgRNAs or a control cell line with a non-targeting (NT) sgRNA were infected with NL4-3 virus or NL4-3 delVpu virus. The supernatants containing newly released viruses were concentrated and analyzed by Western blotting. The cell lysates of the producer cells were also analyzed by Western blotting. c Quantification of the band intensity of gp41 in b normalized to the intensity of <t>p24.</t> The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. d Virions harvested from infection experiments in b were normalized by p24 levels measured with <t>ELISA</t> and used to infect TZM-bl cells. The infectivity was measured by luciferase assays. The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. e Correlation analysis between c and d . f, g Virions from PSGL-1 knockout or control Jurkat cell lines were collected after 5 days post infection with NL4-3 and pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in f . Scale bar: 100 nm. Quantification of STORM images results were showed in g . The ratios between the average values of two groups and the p values were shown. h, i Virions from producer 293 T cells transfected with PSGL-1, PSGL-1 delCD, or an empty vector were pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in h . Scale bar: 100 nm. Quantification of STORM images were showed in i . The ratios between the average values of two groups and the p values were shown. j, k Concentrated virions were analyzed by cryo-EM analysis. Representative images were shown in j and quantification of images of virions shown in k . Scale bar: 100 nm.
    Commercial P24 Elisa Kit, supplied by Sino Biological, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/commercial p24 elisa kit/product/Sino Biological
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    96
    Sino Biological hiv p24
    Characterization of SARS-CoV-2 Spike pseudovirus. A. Incorporation of SARS-CoV-2 S protein into pseudoviruses. VSV-G and S pseudoviruses were pelleted as described in Materials and Methods and subjected to SDS-PAGE, immunoblotted with anti-Spike and <t>HIV</t> <t>p24</t> as control. B. S pseudovirus transduction of 293T, 293T-ACE2, Vero E6 and Huh 7 cells. 293T, 293T-ACE2, Vero E6 and Huh 7 cells were mock infected or infected with S pseudovirus, VSV-G pseudovirus expressing NanoLuc luciferase. At 48 h post transduction, luciferase activity was measured. Values are mean ± SD and are representative of three independent experiments. C. 293T, 293T-ACE2, Vero E6 and Huh 7 cells transduced with S pseudovirus expressing GFP. Bright field was included to show the presence of cells. Bar, 20μm.
    Hiv P24, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv p24/product/Sino Biological
    Average 96 stars, based on 1 article reviews
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    Image Search Results


    NSC- and A549-derived exosomes significantly enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P

    Journal: International Journal of Nanomedicine

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells

    doi: 10.2147/IJN.S132762

    Figure Lengend Snippet: NSC- and A549-derived exosomes significantly enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) NSC-derived exosomes (0.1 μg) or ( C ) A549-dervied exosomes (0.1 μg). ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) NSC-derived exosomes (0.1 μg) or ( D ) A549-derived exosomes (0.1 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA * P

    Article Snippet: Blocking of HIV-1 infection A protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control.

    Techniques: Derivative Assay, Expressing, Luciferase, Activity Assay, Infection

    Breast milk- and plasma-derived exosomes enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) breast milk-derived exosomes (0.035 μg) or ( C ) plasma-derived exosomes (0.05 μg). Virus entry was evaluated in the presence of exosomes and anti-TIM-4 antibodies. ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) breast milk-derived exosomes (0.035 μg) or ( D ) plasma-derived exosomes (0.05 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA. ** P

    Journal: International Journal of Nanomedicine

    Article Title: Role of TIM-4 in exosome-dependent entry of HIV-1 into human immune cells

    doi: 10.2147/IJN.S132762

    Figure Lengend Snippet: Breast milk- and plasma-derived exosomes enhance HIV-1 entry into human immune cell lines. Notes: ( A , C ) YU-2 virus entry into A3R5.7 cells was evaluated in the presence or absence of ( A ) breast milk-derived exosomes (0.035 μg) or ( C ) plasma-derived exosomes (0.05 μg). Virus entry was evaluated in the presence of exosomes and anti-TIM-4 antibodies. ( B , D ) The differentiated THP2574 cell line was used for entry experiments with YU-2 in the presence or absence of ( B ) breast milk-derived exosomes (0.035 μg) or ( D ) plasma-derived exosomes (0.05 μg). Virus entry was also evaluated in the presence of exosomes and anti-TIM-4 antibody. Viral gene expression in all control and treatment groups was assessed by Renilla luciferase activity at 72 h post-infection. Data represent 12 independent experiments. Significant differences between treatment groups were determined by one-way ANOVA. ** P

    Article Snippet: Blocking of HIV-1 infection A protocol similar to HIV-1 infection was performed but with addition of 0.2 μg/well anti-mouse TIM-4 or anti-human TIM-4 (Sino Biological, Inc.) to the YU-2/exosome/cell incubation or with YU-2 only as a control.

    Techniques: Derivative Assay, Expressing, Luciferase, Activity Assay, Infection

    Effects of GPI-anchored antibodies on HIV-1 Env processing. A The expression levels of gp160, gp120, gp41, P24, and β-actin in HEK293T cells coexpressing GPI-m36.4 or GPI-FluIgG03 and NL4-3 (left panel) or THRO.c/2626 (right panel) were detected by Western blotting. For clarity, images were spliced and grouped, and the original images are provided in the Supplementary materials . B The ratio of gp120 to gp160 in cell lysates was determined by quantifying the corresponding band intensities with ImageJ. The data shown were derived from three independent experiments, and error bars indicate standard deviations. Statistical comparisons were conducted by ANOVA (** P

    Journal: Cellular and Molecular Immunology

    Article Title: Generation of HIV-resistant cells with a single-domain antibody: implications for HIV-1 gene therapy

    doi: 10.1038/s41423-020-00627-y

    Figure Lengend Snippet: Effects of GPI-anchored antibodies on HIV-1 Env processing. A The expression levels of gp160, gp120, gp41, P24, and β-actin in HEK293T cells coexpressing GPI-m36.4 or GPI-FluIgG03 and NL4-3 (left panel) or THRO.c/2626 (right panel) were detected by Western blotting. For clarity, images were spliced and grouped, and the original images are provided in the Supplementary materials . B The ratio of gp120 to gp160 in cell lysates was determined by quantifying the corresponding band intensities with ImageJ. The data shown were derived from three independent experiments, and error bars indicate standard deviations. Statistical comparisons were conducted by ANOVA (** P

    Article Snippet: The cell proteins were separated by SDS-PAGE and transferred to a nitrocellulose membrane, which was then blocked with a 5% nonfat dry milk solution in TBS-Tween 20 at room temperature for 1 h. The membrane was incubated overnight at 4 °C with a rabbit anti-gp120 polyclonal antibody (SinoBiological, Beijing, China), the human anti-gp41 monoclonal antibody 10E8, a mouse anti-P24 antibody (Abcam), or a mouse anti-β actin antibody (Sigma).

    Techniques: Expressing, Western Blot, Derivative Assay

    PSGL-1 inhibits Env incorporation in virions and viral entry. a Virions harvested from producer 293 T cells transfected with two different doses of PSGL-1 or an empty vector were pelleted through 20% sucrose cushion were analyzed by Western blotting together with the cell lysates. b PSGL-1-knockout Jurkat cells lines with two different sgRNAs or a control cell line with a non-targeting (NT) sgRNA were infected with NL4-3 virus or NL4-3 delVpu virus. The supernatants containing newly released viruses were concentrated and analyzed by Western blotting. The cell lysates of the producer cells were also analyzed by Western blotting. c Quantification of the band intensity of gp41 in b normalized to the intensity of p24. The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. d Virions harvested from infection experiments in b were normalized by p24 levels measured with ELISA and used to infect TZM-bl cells. The infectivity was measured by luciferase assays. The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. e Correlation analysis between c and d . f, g Virions from PSGL-1 knockout or control Jurkat cell lines were collected after 5 days post infection with NL4-3 and pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in f . Scale bar: 100 nm. Quantification of STORM images results were showed in g . The ratios between the average values of two groups and the p values were shown. h, i Virions from producer 293 T cells transfected with PSGL-1, PSGL-1 delCD, or an empty vector were pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in h . Scale bar: 100 nm. Quantification of STORM images were showed in i . The ratios between the average values of two groups and the p values were shown. j, k Concentrated virions were analyzed by cryo-EM analysis. Representative images were shown in j and quantification of images of virions shown in k . Scale bar: 100 nm.

    Journal: Cell Discovery

    Article Title: PSGL-1 inhibits HIV-1 infection by restricting actin dynamics and sequestering HIV envelope proteins

    doi: 10.1038/s41421-020-0184-9

    Figure Lengend Snippet: PSGL-1 inhibits Env incorporation in virions and viral entry. a Virions harvested from producer 293 T cells transfected with two different doses of PSGL-1 or an empty vector were pelleted through 20% sucrose cushion were analyzed by Western blotting together with the cell lysates. b PSGL-1-knockout Jurkat cells lines with two different sgRNAs or a control cell line with a non-targeting (NT) sgRNA were infected with NL4-3 virus or NL4-3 delVpu virus. The supernatants containing newly released viruses were concentrated and analyzed by Western blotting. The cell lysates of the producer cells were also analyzed by Western blotting. c Quantification of the band intensity of gp41 in b normalized to the intensity of p24. The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. d Virions harvested from infection experiments in b were normalized by p24 levels measured with ELISA and used to infect TZM-bl cells. The infectivity was measured by luciferase assays. The fold of change between NL4-3 virus and NL4-3 delVpu virus is shown for each cell group. e Correlation analysis between c and d . f, g Virions from PSGL-1 knockout or control Jurkat cell lines were collected after 5 days post infection with NL4-3 and pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in f . Scale bar: 100 nm. Quantification of STORM images results were showed in g . The ratios between the average values of two groups and the p values were shown. h, i Virions from producer 293 T cells transfected with PSGL-1, PSGL-1 delCD, or an empty vector were pelleted through 20% sucrose cushion were fixed and stained for STORM imaging. Representative images are shown in h . Scale bar: 100 nm. Quantification of STORM images were showed in i . The ratios between the average values of two groups and the p values were shown. j, k Concentrated virions were analyzed by cryo-EM analysis. Representative images were shown in j and quantification of images of virions shown in k . Scale bar: 100 nm.

    Article Snippet: Two days post transfection, the supernatant was collected for p24 measurement using a commercial p24 ELISA kit (Sinobiological).

    Techniques: Transfection, Plasmid Preparation, Western Blot, Knock-Out, Infection, Enzyme-linked Immunosorbent Assay, Luciferase, Staining, Imaging

    PSGL-1 increases F-actin intensity inside HIV-1 virions and affects virion infectivity. a TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 delCD. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were quantitated with luciferase assay. n = 3. b, c Jurkat cells were infected with Vpr-BlaM virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 delCD. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were measured by FACS. n = 3. d NL4-3 virions packaged from 293 T cells with or without PSGL-1 overexpression were preincubated with indicated doses of F-actin inhibitors, latrunculin A, or cytochalasin D. TZM-bl cells were infected with treated virions for 48 h before the infection units were measured with luciferase assay. n = 3. e Vpr-BlaM containing virions generated from 293 T cells with or without PSGL-1 overexpression were preincubated with latrunculin A or cytochalasin D at indicated concentrations for 2 h. Jurkat cells were infected with the treated virions for 2 h and incubated with β-lactamase substrate overnight before being fixed and analyzed by FACS. n = 3. f Virions harvested from producer 293 T cells transfected with PSGL-1 or PSGL-1 T393A or an empty vector were pelleted through 20% sucrose cushion, fixed by 4% PFA and stained with antibodies and phalloidin before STORM imaging. Scale bar: 100 nm. g Quantification of the intensities of F-actin or PSGL-1 staining in f shows that PSGL-1 but not PSGL-1 T393A significantly stabilizes F-actin in virions. h Virions containing PSGL-1 or PSGL-1 T393A were pelleted through 20% sucrose cushion before being sedimented for fractions containing G-actin or F-actin and analyzed by Western blotting. i TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 T393A. Empty vector DNA was used to normalize the total DNA transfected. The infection rates were quantitated by luciferase assay. n = 3. j Vpr-BlaM containing virions generated from 293 T cells transfected with pNL4-3, Vpr-BlaM plasmid, and different amounts of PSGL-1 or PSGL-1 T393A plasmids. Empty vector was used to normalize the total DNA transfected. The viruses harvested were normalized by p24 ELISA and used to infect Jurkat cells for 2 h. The cells were then incubated with β-lactamase substrate overnight before being fixed and analyzed by FACS. n = 3.

    Journal: Cell Discovery

    Article Title: PSGL-1 inhibits HIV-1 infection by restricting actin dynamics and sequestering HIV envelope proteins

    doi: 10.1038/s41421-020-0184-9

    Figure Lengend Snippet: PSGL-1 increases F-actin intensity inside HIV-1 virions and affects virion infectivity. a TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 delCD. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were quantitated with luciferase assay. n = 3. b, c Jurkat cells were infected with Vpr-BlaM virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 delCD. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were measured by FACS. n = 3. d NL4-3 virions packaged from 293 T cells with or without PSGL-1 overexpression were preincubated with indicated doses of F-actin inhibitors, latrunculin A, or cytochalasin D. TZM-bl cells were infected with treated virions for 48 h before the infection units were measured with luciferase assay. n = 3. e Vpr-BlaM containing virions generated from 293 T cells with or without PSGL-1 overexpression were preincubated with latrunculin A or cytochalasin D at indicated concentrations for 2 h. Jurkat cells were infected with the treated virions for 2 h and incubated with β-lactamase substrate overnight before being fixed and analyzed by FACS. n = 3. f Virions harvested from producer 293 T cells transfected with PSGL-1 or PSGL-1 T393A or an empty vector were pelleted through 20% sucrose cushion, fixed by 4% PFA and stained with antibodies and phalloidin before STORM imaging. Scale bar: 100 nm. g Quantification of the intensities of F-actin or PSGL-1 staining in f shows that PSGL-1 but not PSGL-1 T393A significantly stabilizes F-actin in virions. h Virions containing PSGL-1 or PSGL-1 T393A were pelleted through 20% sucrose cushion before being sedimented for fractions containing G-actin or F-actin and analyzed by Western blotting. i TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1 and PSGL-1 T393A. Empty vector DNA was used to normalize the total DNA transfected. The infection rates were quantitated by luciferase assay. n = 3. j Vpr-BlaM containing virions generated from 293 T cells transfected with pNL4-3, Vpr-BlaM plasmid, and different amounts of PSGL-1 or PSGL-1 T393A plasmids. Empty vector was used to normalize the total DNA transfected. The viruses harvested were normalized by p24 ELISA and used to infect Jurkat cells for 2 h. The cells were then incubated with β-lactamase substrate overnight before being fixed and analyzed by FACS. n = 3.

    Article Snippet: Two days post transfection, the supernatant was collected for p24 measurement using a commercial p24 ELISA kit (Sinobiological).

    Techniques: Infection, Transfection, Expressing, Plasmid Preparation, Enzyme-linked Immunosorbent Assay, Luciferase, FACS, Over Expression, Generated, Incubation, Staining, Imaging, Western Blot

    PSGL-1 stabilizes cellular F-actin to restrict HIV infection. a Immunofluorescence staining of PSGL-1 in HeLa-based MAGI cells overexpressing PSGL-1 using anti-PSGL-1 antibody. Upper panel: PSGL-1 (red) colocalizes with LifeAct-GFP (green), which binds F-actin; Lower panel: PSGL-1 (red) colocalizes with phalloidin (green). Scale bar: 10 μm. b, c Immunofluorescence staining of PSGL-1 using anti-PSGL-1 antibody (green) and phalloidin (red) in Jurkat T cells overexpressing luciferase or PSGL-1. Scale bar: 5 μm. The phalloidin intensity of cells in each group is shown in c . d Jurkat T cells overexpressing luciferase, PSGL-1 or PSGL-1 CD (cytoplasmic domain) alone were stained with phalloidin and analyzed with FACS. Relative MFIs were normalized to luciferase group. MFI mean fluorescence intensity. n = 3. e, f Activated primary CD4 + T cells were treated with IFN-γ or mock-treated for 12 h before being electroporated with two different siRNAs targeting PSGL-1 or non-targeting control siRNA (siNT) for 48 h. The cells were then either fixed for phalloidin staining and FACS quantification ( f ) or infected with HIV-1 NL4-3 for 72 h before the supernatant being collected for p24 ELISA ( e ). n = 3 for e, f . g Correlation between cellular F-actin intensities and HIV-1 infection rates in e, f . h, i Activated primary CD4 + T cells from two healthy donors were incubated with PSGL-1 antibody or IgG for 2 h before being fixed for phalloidin staining ( h ) or infected with HIV-1 NL4-3 for 72 h before the supernatant being collected for p24 measurement ( i ). n = 3.

    Journal: Cell Discovery

    Article Title: PSGL-1 inhibits HIV-1 infection by restricting actin dynamics and sequestering HIV envelope proteins

    doi: 10.1038/s41421-020-0184-9

    Figure Lengend Snippet: PSGL-1 stabilizes cellular F-actin to restrict HIV infection. a Immunofluorescence staining of PSGL-1 in HeLa-based MAGI cells overexpressing PSGL-1 using anti-PSGL-1 antibody. Upper panel: PSGL-1 (red) colocalizes with LifeAct-GFP (green), which binds F-actin; Lower panel: PSGL-1 (red) colocalizes with phalloidin (green). Scale bar: 10 μm. b, c Immunofluorescence staining of PSGL-1 using anti-PSGL-1 antibody (green) and phalloidin (red) in Jurkat T cells overexpressing luciferase or PSGL-1. Scale bar: 5 μm. The phalloidin intensity of cells in each group is shown in c . d Jurkat T cells overexpressing luciferase, PSGL-1 or PSGL-1 CD (cytoplasmic domain) alone were stained with phalloidin and analyzed with FACS. Relative MFIs were normalized to luciferase group. MFI mean fluorescence intensity. n = 3. e, f Activated primary CD4 + T cells were treated with IFN-γ or mock-treated for 12 h before being electroporated with two different siRNAs targeting PSGL-1 or non-targeting control siRNA (siNT) for 48 h. The cells were then either fixed for phalloidin staining and FACS quantification ( f ) or infected with HIV-1 NL4-3 for 72 h before the supernatant being collected for p24 ELISA ( e ). n = 3 for e, f . g Correlation between cellular F-actin intensities and HIV-1 infection rates in e, f . h, i Activated primary CD4 + T cells from two healthy donors were incubated with PSGL-1 antibody or IgG for 2 h before being fixed for phalloidin staining ( h ) or infected with HIV-1 NL4-3 for 72 h before the supernatant being collected for p24 measurement ( i ). n = 3.

    Article Snippet: Two days post transfection, the supernatant was collected for p24 measurement using a commercial p24 ELISA kit (Sinobiological).

    Techniques: Infection, Immunofluorescence, Staining, Luciferase, FACS, Fluorescence, Enzyme-linked Immunosorbent Assay, Incubation

    PSGL-1 binds HIV Env and sequesters Env at the plasma membrane. a 293 T cells were separately transfected with pNL4-3 proviral plasmid or plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA, or an empty vector. One day after the transfection, PSGL-1 or gp41 were immunoprecipitated with protein A agarose beads with anti-PSGL-1 antibody or anti-gp41 antibody respectively and then the beads were mixed with cell lysates from transfection of the other protein in the IP. The cell lysates and the precipitated proteins were analyzed by Western blotting. b 293 T cells transfected with pNL4-3 and PSGL-1 or PSGL-1 delCD or PSGL-1 LL/AA or an empty vector were fixed and stained with anti-gp41 (red), anti-PSGL-1 (green) antibodies and DAPI (blue), and quantification of gp41 cellular localization of each sample is shown in c . Scale bar: 5 μm. n = 40. d Virions harvested from producer 293 T cells transfected with PSGL-1 or PSGL-1 delCD or PSGL-1 LL/AA or an empty vector were pelleted through 20% sucrose cushion were analyzed by Western blotting. e TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were quantitated with the luciferase assay. n = 3. f Endocytosis of Env (gp160) protein in 293 T cells as visualized by pulse labeling of FAP-tag in the Env protein. 293 T cells were transfected with plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA and then stained with the fluorogen MG-11p (red) for 5 min. Cells were fixed after a 20-min chase period and stained with an anti-PSGL-1 (green) antibody and DAPI (blue). Scale bar: 5 μm. g Model of the mechanisms of the anti-HIV activity of PSGL-1. In the early stage of HIV life cycle, PSGL-1 restricts cortical actin filament to inhibit HIV reverse transcription. In the late stage of the life cycle, PSGL-1 prevents envelope incorporation into the nascent virions and blocks viral infectivity. PSGL-1 also stabilize F-actin inside of nascent virions to affect their infectivity.

    Journal: Cell Discovery

    Article Title: PSGL-1 inhibits HIV-1 infection by restricting actin dynamics and sequestering HIV envelope proteins

    doi: 10.1038/s41421-020-0184-9

    Figure Lengend Snippet: PSGL-1 binds HIV Env and sequesters Env at the plasma membrane. a 293 T cells were separately transfected with pNL4-3 proviral plasmid or plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA, or an empty vector. One day after the transfection, PSGL-1 or gp41 were immunoprecipitated with protein A agarose beads with anti-PSGL-1 antibody or anti-gp41 antibody respectively and then the beads were mixed with cell lysates from transfection of the other protein in the IP. The cell lysates and the precipitated proteins were analyzed by Western blotting. b 293 T cells transfected with pNL4-3 and PSGL-1 or PSGL-1 delCD or PSGL-1 LL/AA or an empty vector were fixed and stained with anti-gp41 (red), anti-PSGL-1 (green) antibodies and DAPI (blue), and quantification of gp41 cellular localization of each sample is shown in c . Scale bar: 5 μm. n = 40. d Virions harvested from producer 293 T cells transfected with PSGL-1 or PSGL-1 delCD or PSGL-1 LL/AA or an empty vector were pelleted through 20% sucrose cushion were analyzed by Western blotting. e TZM-bl cells were infected with virions harvested from 293 T cells transfected with pNL4-3 and different amounts of plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA. Empty vector was used to normalize the total transfected DNA. The virions were normalized by p24 ELISA. The infection rates were quantitated with the luciferase assay. n = 3. f Endocytosis of Env (gp160) protein in 293 T cells as visualized by pulse labeling of FAP-tag in the Env protein. 293 T cells were transfected with plasmids expressing PSGL-1, PSGL-1 delCD, PSGL-1 LL/AA and then stained with the fluorogen MG-11p (red) for 5 min. Cells were fixed after a 20-min chase period and stained with an anti-PSGL-1 (green) antibody and DAPI (blue). Scale bar: 5 μm. g Model of the mechanisms of the anti-HIV activity of PSGL-1. In the early stage of HIV life cycle, PSGL-1 restricts cortical actin filament to inhibit HIV reverse transcription. In the late stage of the life cycle, PSGL-1 prevents envelope incorporation into the nascent virions and blocks viral infectivity. PSGL-1 also stabilize F-actin inside of nascent virions to affect their infectivity.

    Article Snippet: Two days post transfection, the supernatant was collected for p24 measurement using a commercial p24 ELISA kit (Sinobiological).

    Techniques: Transfection, Plasmid Preparation, Expressing, Immunoprecipitation, Western Blot, Staining, Infection, Enzyme-linked Immunosorbent Assay, Luciferase, Labeling, Activity Assay

    Characterization of SARS-CoV-2 Spike pseudovirus. A. Incorporation of SARS-CoV-2 S protein into pseudoviruses. VSV-G and S pseudoviruses were pelleted as described in Materials and Methods and subjected to SDS-PAGE, immunoblotted with anti-Spike and HIV p24 as control. B. S pseudovirus transduction of 293T, 293T-ACE2, Vero E6 and Huh 7 cells. 293T, 293T-ACE2, Vero E6 and Huh 7 cells were mock infected or infected with S pseudovirus, VSV-G pseudovirus expressing NanoLuc luciferase. At 48 h post transduction, luciferase activity was measured. Values are mean ± SD and are representative of three independent experiments. C. 293T, 293T-ACE2, Vero E6 and Huh 7 cells transduced with S pseudovirus expressing GFP. Bright field was included to show the presence of cells. Bar, 20μm.

    Journal: bioRxiv

    Article Title: Cytoplasmic domain and enzymatic activity of ACE2 is not required for PI4KB dependent endocytosis entry of SARS-CoV-2 into host cells

    doi: 10.1101/2021.03.01.433503

    Figure Lengend Snippet: Characterization of SARS-CoV-2 Spike pseudovirus. A. Incorporation of SARS-CoV-2 S protein into pseudoviruses. VSV-G and S pseudoviruses were pelleted as described in Materials and Methods and subjected to SDS-PAGE, immunoblotted with anti-Spike and HIV p24 as control. B. S pseudovirus transduction of 293T, 293T-ACE2, Vero E6 and Huh 7 cells. 293T, 293T-ACE2, Vero E6 and Huh 7 cells were mock infected or infected with S pseudovirus, VSV-G pseudovirus expressing NanoLuc luciferase. At 48 h post transduction, luciferase activity was measured. Values are mean ± SD and are representative of three independent experiments. C. 293T, 293T-ACE2, Vero E6 and Huh 7 cells transduced with S pseudovirus expressing GFP. Bright field was included to show the presence of cells. Bar, 20μm.

    Article Snippet: Antibodies and reagents Antibodies used in this study include: SARS-CoV-2 spike (GeneTex, Taiwan), HA tag, EEA1 (Cell Signaling Technology, Danvers, MA), HIV-p24 (SinoBiological, Beijing, China), β-actin (ABclonal, Woburn, MA), CHC (Santa Cruz Biotechnology, Dallas, TX), CAV1(Proteintech, Chicago, IL) and ACE2 (Abways Technology, Shanghai, China).

    Techniques: SDS Page, Transduction, Infection, Expressing, Luciferase, Activity Assay