sars cov sars cov 2 nucleocapsid antibody mouse mab  (Sino Biological)


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    Name:
    SARS CoV SARS CoV 2 Nucleocapsid Antibody Mouse MAb
    Description:
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant SARS CoV Nucleoprotein NP Catalog 40143 V08B NP 828858 1 Met1 Ala422 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
    Catalog Number:
    40143-MM05
    Price:
    None
    Category:
    Primary Antibody
    Reactivity:
    SARS
    Applications:
    WB,ELISA
    Immunogen:
    Recombinant SARS-CoV Nucleoprotein / NP Protein (Catalog#40143-V08B)
    Product Aliases:
    Anti-coronavirus NP Antibody, Anti-coronavirus Nucleocapsid Antibody, Anti-coronavirus Nucleoprotein Antibody, Anti-cov np Antibody, Anti-ncov NP Antibody, Anti-novel coronavirus NP Antibody, Anti-novel coronavirus Nucleocapsid Antibody, Anti-novel coronavirus Nucleoprotein Antibody, Anti-NP Antibody, Anti-Nucleocapsid Antibody, Anti-Nucleoprotein Antibody
    Antibody Type:
    MAb
    Host:
    Mouse
    Isotype:
    Mouse IgG1
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    Structured Review

    Sino Biological sars cov sars cov 2 nucleocapsid antibody mouse mab
    Counter screening of confirmed HCoV 229E hits with <t>SARS-CoV-2.</t> Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.
    This antibody was produced from a hybridoma resulting from the fusion of a mouse myeloma with B cells obtained from a mouse immunized with purified recombinant SARS CoV Nucleoprotein NP Catalog 40143 V08B NP 828858 1 Met1 Ala422 The IgG fraction of the cell culture supernatant was purified by Protein A affinity chromatography
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    sars cov sars cov 2 nucleocapsid antibody mouse mab - by Bioz Stars, 2021-09
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    Images

    1) Product Images from "Repurposing screen highlights broad-spectrum coronavirus antivirals and their host targets"

    Article Title: Repurposing screen highlights broad-spectrum coronavirus antivirals and their host targets

    Journal: bioRxiv

    doi: 10.1101/2021.07.14.452343

    Counter screening of confirmed HCoV 229E hits with SARS-CoV-2. Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.
    Figure Legend Snippet: Counter screening of confirmed HCoV 229E hits with SARS-CoV-2. Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.

    Techniques Used: Infection, Concentration Assay, Staining, Imaging, Expressing, Quantitative RT-PCR, Immunofluorescence, Activity Assay

    Results of ReFRAME library screening against the alphacoronavirus HCoV 229E. (A) Results of the dual luciferase-based HCoV 229E/R-Luc screening of the ReFRAME library in Huh-7.5/F-Luc cells. Renilla luciferase (R-Luc) activity is proportional to virus infection, whereas firefly luciferase (F-Luc) activity corresponds to cell numbers and viability. R-Luc and F-Luc values detected in DMSO-treated HCoV 229E infected cells were set to 100% and corrected for background values in case of R-Luc. (B) Stage of development of confirmed HCoV 229E hit compounds and information about previous screens by others. (C) Fifty-nine distinct host targets are annotated to these hits. The number before the dash indicates the number of antiviral hits with this target annotation, the number after the dash is equivalent to the total number of molecules within this target category in the ReFRAME library. (D) Overlap between this present ReFRAME library screening with HCoV 229E confirmed hits of the previous SARS-CoV-2 screening by Riva at al. ( Riva et al., 2020 ). Numbers indicate the IC50 ranks of the respective studies. (E) Dose-dependent antiviral activity of top ranked confirmed hits of Riva et al. against HCoV 229E. Dose-dependent antiviral activity of CDK inhibitors (F) and three inflammatory modulators (G) against HCoV 229E. Numbers before compound name indicate IC50-rank in the HCoV 229E screening. One experiment with means of two independent luciferase measurements normalized to DMSO control is given.
    Figure Legend Snippet: Results of ReFRAME library screening against the alphacoronavirus HCoV 229E. (A) Results of the dual luciferase-based HCoV 229E/R-Luc screening of the ReFRAME library in Huh-7.5/F-Luc cells. Renilla luciferase (R-Luc) activity is proportional to virus infection, whereas firefly luciferase (F-Luc) activity corresponds to cell numbers and viability. R-Luc and F-Luc values detected in DMSO-treated HCoV 229E infected cells were set to 100% and corrected for background values in case of R-Luc. (B) Stage of development of confirmed HCoV 229E hit compounds and information about previous screens by others. (C) Fifty-nine distinct host targets are annotated to these hits. The number before the dash indicates the number of antiviral hits with this target annotation, the number after the dash is equivalent to the total number of molecules within this target category in the ReFRAME library. (D) Overlap between this present ReFRAME library screening with HCoV 229E confirmed hits of the previous SARS-CoV-2 screening by Riva at al. ( Riva et al., 2020 ). Numbers indicate the IC50 ranks of the respective studies. (E) Dose-dependent antiviral activity of top ranked confirmed hits of Riva et al. against HCoV 229E. Dose-dependent antiviral activity of CDK inhibitors (F) and three inflammatory modulators (G) against HCoV 229E. Numbers before compound name indicate IC50-rank in the HCoV 229E screening. One experiment with means of two independent luciferase measurements normalized to DMSO control is given.

    Techniques Used: Library Screening, Luciferase, Activity Assay, Infection

    Validation of candidates with broad-spectrum coronavirus activity in well differentiated primary human lung cells. Primary cells from three independent donors were differentiated and grown as pseudostratified epithelium in air liquid interface culture. Cells were pre-treated for 2h with final concentrations of 1 µM (bright blue) or 5 µM (dark blue) of compounds from the basolateral side prior to inoculation with SARS-CoV-2 in the presence of compounds (apical and basal treatment). Infection efficiency was determined 72h post inoculation by qRT-PCR of cell extracts (A) and culture fluids (B). (C) Cell viability was assessed by quantification of GAPDH mRNA. Ct values > 35 were excluded from the analysis. Mean values (bar) and individual values (points) of three donors are given. Analysis was performed in two independent experiments (as indicated by the separation). In both experiments, primary cells from three different donors were used. Virus stocks differed in titer between these experiments (see Material and Methods).
    Figure Legend Snippet: Validation of candidates with broad-spectrum coronavirus activity in well differentiated primary human lung cells. Primary cells from three independent donors were differentiated and grown as pseudostratified epithelium in air liquid interface culture. Cells were pre-treated for 2h with final concentrations of 1 µM (bright blue) or 5 µM (dark blue) of compounds from the basolateral side prior to inoculation with SARS-CoV-2 in the presence of compounds (apical and basal treatment). Infection efficiency was determined 72h post inoculation by qRT-PCR of cell extracts (A) and culture fluids (B). (C) Cell viability was assessed by quantification of GAPDH mRNA. Ct values > 35 were excluded from the analysis. Mean values (bar) and individual values (points) of three donors are given. Analysis was performed in two independent experiments (as indicated by the separation). In both experiments, primary cells from three different donors were used. Virus stocks differed in titer between these experiments (see Material and Methods).

    Techniques Used: Activity Assay, Infection, Quantitative RT-PCR

    2) Product Images from "Susceptibility of rabbits to SARS-CoV-2"

    Article Title: Susceptibility of rabbits to SARS-CoV-2

    Journal: bioRxiv

    doi: 10.1101/2020.08.27.263988

    Rabbit ACE2 mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authen tic virus (B) infection of Cos-7 cells expressing ACE2 of various species. Infectivity was quantified by staining live virus cells with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2 mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50μm.
    Figure Legend Snippet: Rabbit ACE2 mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authen tic virus (B) infection of Cos-7 cells expressing ACE2 of various species. Infectivity was quantified by staining live virus cells with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2 mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50μm.

    Techniques Used: Infection, Expressing, Staining, Imaging

    3) Product Images from "Coinfection with influenza A virus enhances SARS-CoV-2 infectivity"

    Article Title: Coinfection with influenza A virus enhances SARS-CoV-2 infectivity

    Journal: Cell Research

    doi: 10.1038/s41422-021-00473-1

    IAV and SARS-CoV-2 coinfection induced more severe pathology in infected mice. a Diagram of the experimental procedure. K18-hACE2 transgenic mice were first intranasally infected with 2000 PFU of WSN or PBS on day 0. Two days post-IAV infection, mice were intranasally infected with 3 × 10 5 live SARS-CoV-2 or PBS. On day 4, half of the lung tissues collected from all the mice were homogenized to detect RNA or protein levels. b The body weights and survival were monitored until day 4 (non-IAV treatment group, n = 4; IAV preinfection group, n = 4). The dotted lines indicate the initial weight. The body weights are presented as the mean percentage of weight change ± SD. c The viral genome copy numbers of SARS-CoV-2 N were quantified. Values represent means ± SD of three individual mice. d The relative mRNA levels of IAV NP ( d, left ), SARS-CoV-2 E ( d, middle ) and the N gene ( d, right ) were measured from lung homogenates in the indicated groups and normalized to GAPDH for the individual mouse. The data are expressed as fold changes relative to the non-IAV infection control. Values represent means ± SD of three individual mice. e Histopathologic and immunohistochemical studies were performed with lung slide samples in the indicated groups c – d . * P
    Figure Legend Snippet: IAV and SARS-CoV-2 coinfection induced more severe pathology in infected mice. a Diagram of the experimental procedure. K18-hACE2 transgenic mice were first intranasally infected with 2000 PFU of WSN or PBS on day 0. Two days post-IAV infection, mice were intranasally infected with 3 × 10 5 live SARS-CoV-2 or PBS. On day 4, half of the lung tissues collected from all the mice were homogenized to detect RNA or protein levels. b The body weights and survival were monitored until day 4 (non-IAV treatment group, n = 4; IAV preinfection group, n = 4). The dotted lines indicate the initial weight. The body weights are presented as the mean percentage of weight change ± SD. c The viral genome copy numbers of SARS-CoV-2 N were quantified. Values represent means ± SD of three individual mice. d The relative mRNA levels of IAV NP ( d, left ), SARS-CoV-2 E ( d, middle ) and the N gene ( d, right ) were measured from lung homogenates in the indicated groups and normalized to GAPDH for the individual mouse. The data are expressed as fold changes relative to the non-IAV infection control. Values represent means ± SD of three individual mice. e Histopathologic and immunohistochemical studies were performed with lung slide samples in the indicated groups c – d . * P

    Techniques Used: Infection, Mouse Assay, Transgenic Assay, Immunohistochemistry

    ACE2 is essential for IAV promotion of SARS-CoV-2 infection. a, b A549 cells were mock-infected or infected with WSN at an MOI of 0.1. At 12 h.p.i., total RNA was extracted from cells, and ACE2, TMPRSS2, Furin, and CatL mRNA levels ( a ) or NP, Mx1, and ISG54 mRNA levels ( b ) were evaluated via qRT-PCR using the SYBR green method. The data are expressed as fold changes relative to the mock infections. c A549 cells were infected with WSN at an MOI of 0.1. IAV NP protein (red) and ACE2 (green) were detected with an immunofluorescence assay at 12 h.p.i. Scale bars are shown. A549 ( d ), Calu-3 ( e ), and NHBE ( f ) cells were preinfected with WSN at an MOI of 0.1 for 12 h. Cells were then infected with live SARS-CoV-2 at an MOI of 0.01 for another 48 h. Total RNA was extracted from cells, and ACE2 mRNA was evaluated via qRT-PCR using the SYBR green method. The protein expression levels of ACE2, SARS-CoV-2 N gene, IAV NP, and β-actin were measured via western blotting assay. * means increased exposure to visualize ACE2. ( g ) The relative mRNA levels of ACE2 were measured in lung homogenates from the indicated groups, and the protein expression of IAV NP and ACE2 was detected via western blotting. ( d – g ) The data are expressed as fold changes relative to the non-IAV infection control. ( h – i ) To establish ACE2 knockdown cells, A549 cells were transduced with lentivirus encoding the CRISPR-Cas9 system with two guide RNAs targeting ACE2 (sgRNA1 and sgRNA2) or control guide RNA. Cells were infected with live SARS-CoV-2 at an MOI of 0.01 with or without IAV infection using the same procedure described above. The ACE2 (qRT-PCR) ( h ) and SARS-CoV-2 E gene (Taqman-qRT-PCR) ( i ) mRNA levels were detected. The data are expressed as the fold change relative to the non-IAV infection control. Values represent means ± SD of three independent experiments. * P
    Figure Legend Snippet: ACE2 is essential for IAV promotion of SARS-CoV-2 infection. a, b A549 cells were mock-infected or infected with WSN at an MOI of 0.1. At 12 h.p.i., total RNA was extracted from cells, and ACE2, TMPRSS2, Furin, and CatL mRNA levels ( a ) or NP, Mx1, and ISG54 mRNA levels ( b ) were evaluated via qRT-PCR using the SYBR green method. The data are expressed as fold changes relative to the mock infections. c A549 cells were infected with WSN at an MOI of 0.1. IAV NP protein (red) and ACE2 (green) were detected with an immunofluorescence assay at 12 h.p.i. Scale bars are shown. A549 ( d ), Calu-3 ( e ), and NHBE ( f ) cells were preinfected with WSN at an MOI of 0.1 for 12 h. Cells were then infected with live SARS-CoV-2 at an MOI of 0.01 for another 48 h. Total RNA was extracted from cells, and ACE2 mRNA was evaluated via qRT-PCR using the SYBR green method. The protein expression levels of ACE2, SARS-CoV-2 N gene, IAV NP, and β-actin were measured via western blotting assay. * means increased exposure to visualize ACE2. ( g ) The relative mRNA levels of ACE2 were measured in lung homogenates from the indicated groups, and the protein expression of IAV NP and ACE2 was detected via western blotting. ( d – g ) The data are expressed as fold changes relative to the non-IAV infection control. ( h – i ) To establish ACE2 knockdown cells, A549 cells were transduced with lentivirus encoding the CRISPR-Cas9 system with two guide RNAs targeting ACE2 (sgRNA1 and sgRNA2) or control guide RNA. Cells were infected with live SARS-CoV-2 at an MOI of 0.01 with or without IAV infection using the same procedure described above. The ACE2 (qRT-PCR) ( h ) and SARS-CoV-2 E gene (Taqman-qRT-PCR) ( i ) mRNA levels were detected. The data are expressed as the fold change relative to the non-IAV infection control. Values represent means ± SD of three independent experiments. * P

    Techniques Used: Infection, Quantitative RT-PCR, SYBR Green Assay, Immunofluorescence, Expressing, Western Blot, Transduction, CRISPR

    SARS-CoV-2 infection in response to HRSV, HPIV, or HRV3. a – c A549 cells were preinfected with HRSV, HPIV, or HRV3 at the indicated doses for 12 h. Cells were then infected with pseudo-SARS-CoV-2 for another 24 h followed by measurement of luciferase activity. d – f The propagation of HRSV ( d ), HPIV ( e ), or HRV3 ( f ) in A549 cells was measured by qRT-PCR 12 h.p.i. targeting the individual viral genes respectively. g – i A549 cells were preinfected with HRSV ( g ), HPIV ( h ), or HRV3 ( i ) at the indicated doses for 12 h. Cells were then infected with live SARS-CoV-2 for another 48 h followed by measurement of SARS-CoV-2 E gene copies via Taqman-qRT-PCR. j – l The propagation of HRSV ( j ), HPIV ( k ), or HRV3 ( l ) in A549 cells was measured at 60 h.p.i. by qRT-PCR targeting the individual viral genes respectively. The data are expressed as fold changes relative to the non-preinfection group. Values represent means ± SD of three independent experiments. * P
    Figure Legend Snippet: SARS-CoV-2 infection in response to HRSV, HPIV, or HRV3. a – c A549 cells were preinfected with HRSV, HPIV, or HRV3 at the indicated doses for 12 h. Cells were then infected with pseudo-SARS-CoV-2 for another 24 h followed by measurement of luciferase activity. d – f The propagation of HRSV ( d ), HPIV ( e ), or HRV3 ( f ) in A549 cells was measured by qRT-PCR 12 h.p.i. targeting the individual viral genes respectively. g – i A549 cells were preinfected with HRSV ( g ), HPIV ( h ), or HRV3 ( i ) at the indicated doses for 12 h. Cells were then infected with live SARS-CoV-2 for another 48 h followed by measurement of SARS-CoV-2 E gene copies via Taqman-qRT-PCR. j – l The propagation of HRSV ( j ), HPIV ( k ), or HRV3 ( l ) in A549 cells was measured at 60 h.p.i. by qRT-PCR targeting the individual viral genes respectively. The data are expressed as fold changes relative to the non-preinfection group. Values represent means ± SD of three independent experiments. * P

    Techniques Used: Infection, Luciferase, Activity Assay, Quantitative RT-PCR

    IAV promotes SARS-CoV-2 virus infectivity. a Diagram of the experimental procedure. b A549 cells were infected with A/WSN/33(WSN) at the indicated MOIs. At 6, 12, and 24 h post-IAV infection, cells were infected with pseudo-SARS-CoV-2 for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. P values are from unpaired one-way ANOVA. c A549 cells were infected with WSN at an MOI of 0.1. At 12 h post-IAV infection, cells were infected with VSV-G-Luc for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. d The indicated cells were infected with WSN at an MOI of 0.1. At 12 h post-IAV infection, cells were infected with pseudo-SARS-CoV-2 for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. e Experimental procedure of IAV and live SARS-CoV-2 coinfection. A549 ( f ), Calu-3 ( g ), and NHBE ( h ) cells were preinfected with WSN at an MOI of 0.1 for 12 h. Cells were then infected with live SARS-CoV-2 at an MOI of 0.01 for another 48 h. Total RNA in cell lysates and the supernatants was collected to detect the E and N genes via TaqMan-qRT-PCR. The data are expressed as fold changes in viral RNA levels in IAV preinfected cells relative to the non-IAV infection control. Values represent means ± SD of three independent experiments. * P
    Figure Legend Snippet: IAV promotes SARS-CoV-2 virus infectivity. a Diagram of the experimental procedure. b A549 cells were infected with A/WSN/33(WSN) at the indicated MOIs. At 6, 12, and 24 h post-IAV infection, cells were infected with pseudo-SARS-CoV-2 for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. P values are from unpaired one-way ANOVA. c A549 cells were infected with WSN at an MOI of 0.1. At 12 h post-IAV infection, cells were infected with VSV-G-Luc for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. d The indicated cells were infected with WSN at an MOI of 0.1. At 12 h post-IAV infection, cells were infected with pseudo-SARS-CoV-2 for another 24 h. Luciferase activity was measured to reflect virus entry efficiency. e Experimental procedure of IAV and live SARS-CoV-2 coinfection. A549 ( f ), Calu-3 ( g ), and NHBE ( h ) cells were preinfected with WSN at an MOI of 0.1 for 12 h. Cells were then infected with live SARS-CoV-2 at an MOI of 0.01 for another 48 h. Total RNA in cell lysates and the supernatants was collected to detect the E and N genes via TaqMan-qRT-PCR. The data are expressed as fold changes in viral RNA levels in IAV preinfected cells relative to the non-IAV infection control. Values represent means ± SD of three independent experiments. * P

    Techniques Used: Infection, Luciferase, Activity Assay, Quantitative RT-PCR

    Enhanced SARS-CoV-2 infection is independent of IFN signaling. A549 ( a – c ), Calu-3 ( d – f ), and Huh-7 ( g – i ) cells were pretreated with the indicated doses of IFNα for 12 h. Cells were then infected with pseudo-SARS-CoV-2 for another 24 h, followed by measurement of luciferase activity and the mRNA expression levels of the indicated genes. The mRNA levels are expressed as fold changes relative to nontreated cells. P values are from unpaired one-way ANOVA. j – l WT A549 and IFNAR IFNAR –/– A549 cells were infected with WSN at an MOI of 0.1 for 12 h, and cells were then infected with pseudo-SARS-CoV-2 for another 24 h, followed by measurement of luciferase activity and the mRNA expression levels of the indicated genes. m WT A549 and IFNAR –/– A549 cells were infected with WSN at an MOI of 0.1 for 12 h, and the cells were then infected with live SARS-CoV-2 for another 48 h, followed by measurement of mRNA levels for SARS-CoV-2 E gene (Taqman-qRT-PCR). The data are expressed as fold changes relative to the nontreatment ( b , c, e , f, h, i ) or mock infection ( k, l ) or non-IAV infection ( m ) controls. Values represent means ± SD of three independent experiments. * P
    Figure Legend Snippet: Enhanced SARS-CoV-2 infection is independent of IFN signaling. A549 ( a – c ), Calu-3 ( d – f ), and Huh-7 ( g – i ) cells were pretreated with the indicated doses of IFNα for 12 h. Cells were then infected with pseudo-SARS-CoV-2 for another 24 h, followed by measurement of luciferase activity and the mRNA expression levels of the indicated genes. The mRNA levels are expressed as fold changes relative to nontreated cells. P values are from unpaired one-way ANOVA. j – l WT A549 and IFNAR IFNAR –/– A549 cells were infected with WSN at an MOI of 0.1 for 12 h, and cells were then infected with pseudo-SARS-CoV-2 for another 24 h, followed by measurement of luciferase activity and the mRNA expression levels of the indicated genes. m WT A549 and IFNAR –/– A549 cells were infected with WSN at an MOI of 0.1 for 12 h, and the cells were then infected with live SARS-CoV-2 for another 48 h, followed by measurement of mRNA levels for SARS-CoV-2 E gene (Taqman-qRT-PCR). The data are expressed as fold changes relative to the nontreatment ( b , c, e , f, h, i ) or mock infection ( k, l ) or non-IAV infection ( m ) controls. Values represent means ± SD of three independent experiments. * P

    Techniques Used: Infection, Luciferase, Activity Assay, Expressing, Quantitative RT-PCR

    4) Product Images from "SARS-CoV-2 entry into human airway organoids is serine protease-mediated and facilitated by the multibasic cleavage site"

    Article Title: SARS-CoV-2 entry into human airway organoids is serine protease-mediated and facilitated by the multibasic cleavage site

    Journal: eLife

    doi: 10.7554/eLife.64508

    hAO cultures grown at 2D air–liquid interface are well-differentiated and express ACE2 and TMPRSS2. ( A, B, and C ) Immunofluorescent or immunohistochemistry staining of differentiated airway cultures. Anti-AcTub (green) and anti-FOXJ1 (white) stains ciliated cells ( A ), anti-SCGB1A1 (magenta) stains club cells ( B ), and anti-MUC5AC (yellow) stains goblet cells ( C ). Nuclei are stained with hoechst (blue). ( D and E ) hAO cultures also expressed the SARS-CoV-2 entry receptor ACE2 ( D ) and TMPRSS2 ( E ). Hematoxylin was used as a counterstain in ( D ) and ( E ). Scale bars indicate 20 μm. Representative images are shown from a bronchiolar culture. ACE2, angiotensin-converting enzyme 2; hAO, human airway organoid; TMPRSS2, transmembrane protease serine 2.
    Figure Legend Snippet: hAO cultures grown at 2D air–liquid interface are well-differentiated and express ACE2 and TMPRSS2. ( A, B, and C ) Immunofluorescent or immunohistochemistry staining of differentiated airway cultures. Anti-AcTub (green) and anti-FOXJ1 (white) stains ciliated cells ( A ), anti-SCGB1A1 (magenta) stains club cells ( B ), and anti-MUC5AC (yellow) stains goblet cells ( C ). Nuclei are stained with hoechst (blue). ( D and E ) hAO cultures also expressed the SARS-CoV-2 entry receptor ACE2 ( D ) and TMPRSS2 ( E ). Hematoxylin was used as a counterstain in ( D ) and ( E ). Scale bars indicate 20 μm. Representative images are shown from a bronchiolar culture. ACE2, angiotensin-converting enzyme 2; hAO, human airway organoid; TMPRSS2, transmembrane protease serine 2.

    Techniques Used: Immunohistochemistry, Staining

    SARS-CoV-2 entry and replication are dependent on serine proteases in human airway organoids. ( A and B ) Differentiated bronchiolar ( A ) or bronchial ( B ) hAO cultures were infected at an MOI of 2. Sixteen hours ( A ) or 24 hr ( B ) postinfection they were fixed and stained for viral nucleoprotein (red). Nuclei were stained with hoechst (blue) and actin was stained using phalloidin (white). AcTub stains ciliated cells (green). Scale bars indicate 200 μm in ( A ) and 50 μm in ( B ). Representative images are shown from two independent experiments. ( C–E ) Replication kinetics of SARS-CoV-2 in bronchiolar hAO cultures pretreated with camostat or carrier (DMSO). ( C and D ) TCID50 equivalents (eq.) per mL are shown in culture medium ( C ) and lysed organoids ( D ). Circles indicate DMSO-treated organoids, whereas squares indicate camostat-treated organoids. ( E ) Live virus titers (TCID50/mL) in lysed organoids. Dotted line indicates limit of detection. ( F ) Replication kinetics of SARS-CoV-2 in 2D tracheal air–liquid interface airway cultures pretreated with camostat or carrier (DMSO). TCID50 eq./mL in apical washes are shown. Two-way ANOVA was performed for statistical analysis. Error bars indicate SEM. *p
    Figure Legend Snippet: SARS-CoV-2 entry and replication are dependent on serine proteases in human airway organoids. ( A and B ) Differentiated bronchiolar ( A ) or bronchial ( B ) hAO cultures were infected at an MOI of 2. Sixteen hours ( A ) or 24 hr ( B ) postinfection they were fixed and stained for viral nucleoprotein (red). Nuclei were stained with hoechst (blue) and actin was stained using phalloidin (white). AcTub stains ciliated cells (green). Scale bars indicate 200 μm in ( A ) and 50 μm in ( B ). Representative images are shown from two independent experiments. ( C–E ) Replication kinetics of SARS-CoV-2 in bronchiolar hAO cultures pretreated with camostat or carrier (DMSO). ( C and D ) TCID50 equivalents (eq.) per mL are shown in culture medium ( C ) and lysed organoids ( D ). Circles indicate DMSO-treated organoids, whereas squares indicate camostat-treated organoids. ( E ) Live virus titers (TCID50/mL) in lysed organoids. Dotted line indicates limit of detection. ( F ) Replication kinetics of SARS-CoV-2 in 2D tracheal air–liquid interface airway cultures pretreated with camostat or carrier (DMSO). TCID50 eq./mL in apical washes are shown. Two-way ANOVA was performed for statistical analysis. Error bars indicate SEM. *p

    Techniques Used: Infection, Staining

    The SARS-CoV-2 multibasic cleavage site increases serine protease usage. ( A and B ) SARS-CoV PP and SARS-CoV-2 PP entry route on VeroE6 cells pretreated with a concentration range of camostat ( A ) or E64D ( B ) to inhibit serine proteases and cathepsins, respectively. ( C and D ) SARS-CoV PP and SARS-CoV-2 PP entry route on VeroE6-TMPRSS2 cells pretreated with a concentration range of camostat ( C ) or E64D ( D ) to inhibit serine proteases and cathepsins, respectively. T-test was performed for statistical analysis at the highest concentration. *p
    Figure Legend Snippet: The SARS-CoV-2 multibasic cleavage site increases serine protease usage. ( A and B ) SARS-CoV PP and SARS-CoV-2 PP entry route on VeroE6 cells pretreated with a concentration range of camostat ( A ) or E64D ( B ) to inhibit serine proteases and cathepsins, respectively. ( C and D ) SARS-CoV PP and SARS-CoV-2 PP entry route on VeroE6-TMPRSS2 cells pretreated with a concentration range of camostat ( C ) or E64D ( D ) to inhibit serine proteases and cathepsins, respectively. T-test was performed for statistical analysis at the highest concentration. *p

    Techniques Used: Concentration Assay

    A GFP complementation based assay for assessing coronavirus fusogenicity. ( A ) HEK-293T cells expressing an empty vector or S protein together with GFP-11-tagged beta-actin and a BFP containing a nuclear localization signal were added to cells stably expressing GFP1-10. Fusion of these two cell types allowed GFP complementation in cells expressing a nuclear BFP, facilitating easy quantification of nuclei per syncytial cell. Unfused cells only expressed BFP in the nucleus. Fusion with VeroE6 GFP1-10 cells 18 hr after addition of the fusogenic HEK-293T is shown as an example. ( B–D ) Full well scans of the complemented GFP signal 18 hr after addition of the fusogenic HEK-293T cells to Calu-3 GFP1-10 ( B ), VeroE6 GFP1-10 ( C ), and VeroE6-TMPRSS2 GFP1-10 ( D ) cells are shown. Dashed areas are enlarged next to each well. Scale bars indicate 50 μm. ( E and F ) Fusogenicity of SARS-CoV-2 S and SARS-CoV S was assessed after 18 hr by measuring the sum of all GFP+ pixels per well in VeroE6 cells ( E ) and VeroE6 TMPRSS2 cells ( F ). Statistical analysis was performed by one-way ANOVA on SARS-CoV-2 S-mediated fusion compared with SARS-CoV S. *p
    Figure Legend Snippet: A GFP complementation based assay for assessing coronavirus fusogenicity. ( A ) HEK-293T cells expressing an empty vector or S protein together with GFP-11-tagged beta-actin and a BFP containing a nuclear localization signal were added to cells stably expressing GFP1-10. Fusion of these two cell types allowed GFP complementation in cells expressing a nuclear BFP, facilitating easy quantification of nuclei per syncytial cell. Unfused cells only expressed BFP in the nucleus. Fusion with VeroE6 GFP1-10 cells 18 hr after addition of the fusogenic HEK-293T is shown as an example. ( B–D ) Full well scans of the complemented GFP signal 18 hr after addition of the fusogenic HEK-293T cells to Calu-3 GFP1-10 ( B ), VeroE6 GFP1-10 ( C ), and VeroE6-TMPRSS2 GFP1-10 ( D ) cells are shown. Dashed areas are enlarged next to each well. Scale bars indicate 50 μm. ( E and F ) Fusogenicity of SARS-CoV-2 S and SARS-CoV S was assessed after 18 hr by measuring the sum of all GFP+ pixels per well in VeroE6 cells ( E ) and VeroE6 TMPRSS2 cells ( F ). Statistical analysis was performed by one-way ANOVA on SARS-CoV-2 S-mediated fusion compared with SARS-CoV S. *p

    Techniques Used: Expressing, Plasmid Preparation, Stable Transfection

    5) Product Images from "SARS-CoV-2 induces human plasmacytoid predendritic cell diversification via UNC93B and IRAK4"

    Article Title: SARS-CoV-2 induces human plasmacytoid predendritic cell diversification via UNC93B and IRAK4

    Journal: The Journal of Experimental Medicine

    doi: 10.1084/jem.20201387

    SARS-CoV-2 induces pDC activation in a dose-dependent manner. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 0.04, 0.2, or 1. (A) Dot plot showing pDC activation through the expression of PD-L1 and CD80. Results from one healthy donor representative of n = 3. (B) Quantification of the three populations. Bars represent medians of n = 3 healthy donors from three independent experiments. (C) pDC geometric mean (mean fluorescence intensity [MFI]) of activation markers after 24 h of culture with medium, Flu, or SARS-CoV-2 at an MOI of 1. Histograms represent medians and bars interquartile ranges of n = 5 healthy donors from three independent experiments. *, P
    Figure Legend Snippet: SARS-CoV-2 induces pDC activation in a dose-dependent manner. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 0.04, 0.2, or 1. (A) Dot plot showing pDC activation through the expression of PD-L1 and CD80. Results from one healthy donor representative of n = 3. (B) Quantification of the three populations. Bars represent medians of n = 3 healthy donors from three independent experiments. (C) pDC geometric mean (mean fluorescence intensity [MFI]) of activation markers after 24 h of culture with medium, Flu, or SARS-CoV-2 at an MOI of 1. Histograms represent medians and bars interquartile ranges of n = 5 healthy donors from three independent experiments. *, P

    Techniques Used: Activation Assay, Cell Culture, Expressing, Fluorescence

    HCQ inhibits SARS-CoV-2–induced pDC activation in a dose-dependent manner. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 1 with HCQ or vehicle. (A) Dot plot of pDC diversification with increasing concentration of HCQ or vehicle. Results from one healthy donor representative of n = 3. (B) Dot plot showing OX40L and CD86 in the presence or absence of HCQ. Results from one healthy donor representative of n = 3. (C) Percentage of OX40L high population among pDCs. Bars represent medians of n = 3 healthy donors from three independent experiments. *, P
    Figure Legend Snippet: HCQ inhibits SARS-CoV-2–induced pDC activation in a dose-dependent manner. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 1 with HCQ or vehicle. (A) Dot plot of pDC diversification with increasing concentration of HCQ or vehicle. Results from one healthy donor representative of n = 3. (B) Dot plot showing OX40L and CD86 in the presence or absence of HCQ. Results from one healthy donor representative of n = 3. (C) Percentage of OX40L high population among pDCs. Bars represent medians of n = 3 healthy donors from three independent experiments. *, P

    Techniques Used: Activation Assay, Cell Culture, Concentration Assay

    SARS-CoV-2–activated pDCs produce pro-inflammatory cytokines. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2. (A) Quantification of secreted pro-inflammatory cytokines after 24 h of culture. Bars represent medians of n = 5 healthy donors from three independent experiments. (B) Dot plot showing pDC activation through the expression of PD-L1 and CD80 (upper plots), and intracellular IFN-α and TNF-α in P1, P2, or P3 populations (lower plots). Results from one healthy donor representative of n = 4. (C) Percentages of IFN-α single-positive, IFN-α + TNF-α + double-positive, and TNF-α single-positive cells in P0, P1, P2, or P3 populations. Histograms represent medians and bars interquartile ranges of n = 4 healthy donors from three independent experiments. *, P
    Figure Legend Snippet: SARS-CoV-2–activated pDCs produce pro-inflammatory cytokines. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2. (A) Quantification of secreted pro-inflammatory cytokines after 24 h of culture. Bars represent medians of n = 5 healthy donors from three independent experiments. (B) Dot plot showing pDC activation through the expression of PD-L1 and CD80 (upper plots), and intracellular IFN-α and TNF-α in P1, P2, or P3 populations (lower plots). Results from one healthy donor representative of n = 4. (C) Percentages of IFN-α single-positive, IFN-α + TNF-α + double-positive, and TNF-α single-positive cells in P0, P1, P2, or P3 populations. Histograms represent medians and bars interquartile ranges of n = 4 healthy donors from three independent experiments. *, P

    Techniques Used: Cell Culture, Activation Assay, Expressing

    SARS-CoV-2 induces activation and diversification of tonsilar pDCs. Sorted blood pDCs from healthy donors were cultured for 48 h with medium, SARS-CoV-2, or Flu. (A) Dot plot showing pDC activation and diversification through the expression of PD-L1 and CD80 at 24 h and 48 h. Results from one healthy donor representative of n = 3. (B) Geometric mean (mean fluorescence intensity [MFI]) of pDCs’ activation markers at 48 h. Histograms represent medians and bars interquartile ranges of n = 3 healthy donors from three independent experiments. (C) Quantification of secreted pro-inflammatory cytokines 48 h after culture. Bars represent medians of n = 3 healthy donors from three independent experiments. (D) Dot plot of tonsil pDC activation cultured for 24 h with medium, SARS-CoV-2, or Flu. Results from n = 1 healthy donors. (E) Quantification of pro-inflammatory cytokines of tonsilar pDCs at 24 h. Histograms represent medians of n = 1 healthy donor. *, P
    Figure Legend Snippet: SARS-CoV-2 induces activation and diversification of tonsilar pDCs. Sorted blood pDCs from healthy donors were cultured for 48 h with medium, SARS-CoV-2, or Flu. (A) Dot plot showing pDC activation and diversification through the expression of PD-L1 and CD80 at 24 h and 48 h. Results from one healthy donor representative of n = 3. (B) Geometric mean (mean fluorescence intensity [MFI]) of pDCs’ activation markers at 48 h. Histograms represent medians and bars interquartile ranges of n = 3 healthy donors from three independent experiments. (C) Quantification of secreted pro-inflammatory cytokines 48 h after culture. Bars represent medians of n = 3 healthy donors from three independent experiments. (D) Dot plot of tonsil pDC activation cultured for 24 h with medium, SARS-CoV-2, or Flu. Results from n = 1 healthy donors. (E) Quantification of pro-inflammatory cytokines of tonsilar pDCs at 24 h. Histograms represent medians of n = 1 healthy donor. *, P

    Techniques Used: Activation Assay, Cell Culture, Expressing, Fluorescence

    SARS-CoV-2–induced pDC activation is inhibited by HCQ. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 1 with or without the presence of HCQ. (A) Dot plot showing pDC diversification in P1, P2, and P3 subpopulations in the presence of HCQ. Results from one healthy donor representative of n = 3. (B) Quantification of the three populations. Bars represent medians of n = 3 healthy donors from three independent experiments. (C) Histograms of pDCs’ activation markers. Results from one healthy donor representative of n = 3. (D) Geometric mean (mean fluorescence intensity [MFI]) of activation markers. Histograms represent medians and bars interquartile ranges of n = 3 healthy donors from three independent experiments. (E) Quantification of pro-inflammatory cytokines production. Bars represent medians of n = 3 healthy donors from three independent experiments. *, P
    Figure Legend Snippet: SARS-CoV-2–induced pDC activation is inhibited by HCQ. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, Flu, or SARS-CoV-2 at an MOI of 1 with or without the presence of HCQ. (A) Dot plot showing pDC diversification in P1, P2, and P3 subpopulations in the presence of HCQ. Results from one healthy donor representative of n = 3. (B) Quantification of the three populations. Bars represent medians of n = 3 healthy donors from three independent experiments. (C) Histograms of pDCs’ activation markers. Results from one healthy donor representative of n = 3. (D) Geometric mean (mean fluorescence intensity [MFI]) of activation markers. Histograms represent medians and bars interquartile ranges of n = 3 healthy donors from three independent experiments. (E) Quantification of pro-inflammatory cytokines production. Bars represent medians of n = 3 healthy donors from three independent experiments. *, P

    Techniques Used: Activation Assay, Cell Culture, Fluorescence

    SARS-CoV-2 induces activation and diversification of primary human pDCs. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, SARS-CoV-2, or Flu. (A) Dot plot showing pDC activation and diversification through the expression of PD-L1 and CD80 into P1, P2, and P3 subpopulations. Results from one healthy donor representative of n = 8. (B) Quantification of the three populations. Bars represent medians of n = 8 healthy donors from six independent experiments. (C) Dot plot showing pDC activation from different strains of SARS-CoV-2 isolated from two patients. Results from one healthy donor representative of n = 3. (D) Percentage of live pDCs after 24 h of culture with medium, SARS-CoV-2, or Flu. n = 8 healthy donors from six independent experiments. (E) Histogram of ACE2 expression on pDCs, Vero E6, and 293T-ACE2 (black) compared with the isotype (light gray). Results from one experiment representative of n = 3. (F) Intracellular production of SARS-CoV-2 ribonucleoprotein in Vero E6 and pDCs at 2, 24, or 48 h post-infection (hpi) with SARS-CoV-2. Results from one experiment representative of n = 3. (G) Infectious viral titers in the supernatants of SARS-CoV-2–infected Vero E6 and pDCs at 2, 24, 48, or 72 hpi. Results from one experiment representative of n = 3. **, P
    Figure Legend Snippet: SARS-CoV-2 induces activation and diversification of primary human pDCs. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, SARS-CoV-2, or Flu. (A) Dot plot showing pDC activation and diversification through the expression of PD-L1 and CD80 into P1, P2, and P3 subpopulations. Results from one healthy donor representative of n = 8. (B) Quantification of the three populations. Bars represent medians of n = 8 healthy donors from six independent experiments. (C) Dot plot showing pDC activation from different strains of SARS-CoV-2 isolated from two patients. Results from one healthy donor representative of n = 3. (D) Percentage of live pDCs after 24 h of culture with medium, SARS-CoV-2, or Flu. n = 8 healthy donors from six independent experiments. (E) Histogram of ACE2 expression on pDCs, Vero E6, and 293T-ACE2 (black) compared with the isotype (light gray). Results from one experiment representative of n = 3. (F) Intracellular production of SARS-CoV-2 ribonucleoprotein in Vero E6 and pDCs at 2, 24, or 48 h post-infection (hpi) with SARS-CoV-2. Results from one experiment representative of n = 3. (G) Infectious viral titers in the supernatants of SARS-CoV-2–infected Vero E6 and pDCs at 2, 24, 48, or 72 hpi. Results from one experiment representative of n = 3. **, P

    Techniques Used: Activation Assay, Cell Culture, Expressing, Isolation, Infection

    SARS-CoV-2 induces pDC activation. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, SARS-CoV-2, or Flu. (A) Percentage of pure pDCs among live cells through different sorting strategies. Results from two healthy donors representative of n = 8. (B) P1, P2, and P3 diversification of fresh, fluorescent-sorted pDCs versus frozen, magnetic-sorted pDCs with either SARS-CoV-2 or Flu for 24 h. Results from two healthy donors representative of n = 8. (C) Dot plot of pDC activation with either free SARS-CoV-2 or pDC co-culture with SARS-CoV-2–infected cells. Results from n = 1 healthy donors. (D) Viral RNA copy number of Vero E6 and pDCs 2, 24, and 48 h post-infection (hpi). Results from one experiment representative of n = 3. (E) Intracellular production of the nucleoprotein antigen (N) on pDCs of two healthy donors of n = 3. FSC-A, forward scatter area.
    Figure Legend Snippet: SARS-CoV-2 induces pDC activation. Sorted blood pDCs from healthy donors were cultured for 24 h with medium, SARS-CoV-2, or Flu. (A) Percentage of pure pDCs among live cells through different sorting strategies. Results from two healthy donors representative of n = 8. (B) P1, P2, and P3 diversification of fresh, fluorescent-sorted pDCs versus frozen, magnetic-sorted pDCs with either SARS-CoV-2 or Flu for 24 h. Results from two healthy donors representative of n = 8. (C) Dot plot of pDC activation with either free SARS-CoV-2 or pDC co-culture with SARS-CoV-2–infected cells. Results from n = 1 healthy donors. (D) Viral RNA copy number of Vero E6 and pDCs 2, 24, and 48 h post-infection (hpi). Results from one experiment representative of n = 3. (E) Intracellular production of the nucleoprotein antigen (N) on pDCs of two healthy donors of n = 3. FSC-A, forward scatter area.

    Techniques Used: Activation Assay, Cell Culture, Co-Culture Assay, Infection

    SARS-CoV-2–induced pDC activation requires IRAK4 and UNC93B1. Sorted blood pDCs from mutated patients and healthy donors were cultured for 24 h with either medium or SARS-CoV-2. (A) Dot plot showing pDC diversification in P1, P2, and P3 subpopulations from magnetically sorted blood pDCs from homozygous IRAK4 −/− ( n = 1), UNC93B1 −/− ( n = 2), and TLR3 −/− ( n = 1) donors, and sex- and age-matched healthy donors ( n = 2), were cultured for 24 h with either medium or SARS-CoV-2 at an MOI of 1. (B) Quantification of pro-inflammatory cytokines in the supernatant of activated pDCs for 24 h in response to SARS-CoV-2 challenge. Bars represent medians. HD, healthy donor; ND, not detectable.
    Figure Legend Snippet: SARS-CoV-2–induced pDC activation requires IRAK4 and UNC93B1. Sorted blood pDCs from mutated patients and healthy donors were cultured for 24 h with either medium or SARS-CoV-2. (A) Dot plot showing pDC diversification in P1, P2, and P3 subpopulations from magnetically sorted blood pDCs from homozygous IRAK4 −/− ( n = 1), UNC93B1 −/− ( n = 2), and TLR3 −/− ( n = 1) donors, and sex- and age-matched healthy donors ( n = 2), were cultured for 24 h with either medium or SARS-CoV-2 at an MOI of 1. (B) Quantification of pro-inflammatory cytokines in the supernatant of activated pDCs for 24 h in response to SARS-CoV-2 challenge. Bars represent medians. HD, healthy donor; ND, not detectable.

    Techniques Used: Activation Assay, Cell Culture

    6) Product Images from "Susceptibility of rabbits to SARS-CoV-2"

    Article Title: Susceptibility of rabbits to SARS-CoV-2

    Journal: Emerging Microbes & Infections

    doi: 10.1080/22221751.2020.1868951

    Susceptibility of rabbits to SARS-CoV-2 infection. Infection kinetics of (A) viral RNA and (B) authentic SARS-CoV-2 virus growth curves from rabbits inoculated with 10 6 TCID 50 and followed up for 21 days. (C–E) Viral RNA growth curved in rabbits inoculated with either (C) 10 6 , (D) 10 5 , or (E) 10 4 TCID 50 and followed up for four days post infection. (F) Viral RNA in lung and nasal turbinates of 10 6 TCID 50 infected rabbits, sacrificed after four days. The RNA detection limit was 3.5 × 10 −1 RNA copies/mL, while the live virus detection limit is 12.5 TCID 50 /mL. Error bars depict SEM. n = 3.
    Figure Legend Snippet: Susceptibility of rabbits to SARS-CoV-2 infection. Infection kinetics of (A) viral RNA and (B) authentic SARS-CoV-2 virus growth curves from rabbits inoculated with 10 6 TCID 50 and followed up for 21 days. (C–E) Viral RNA growth curved in rabbits inoculated with either (C) 10 6 , (D) 10 5 , or (E) 10 4 TCID 50 and followed up for four days post infection. (F) Viral RNA in lung and nasal turbinates of 10 6 TCID 50 infected rabbits, sacrificed after four days. The RNA detection limit was 3.5 × 10 −1 RNA copies/mL, while the live virus detection limit is 12.5 TCID 50 /mL. Error bars depict SEM. n = 3.

    Techniques Used: Infection, RNA Detection

    Rabbit ACE2-mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authentic virus (B) infection of Cos-7 cells transfected with ACE2 of various species. Infectivity was quantified by staining live virus with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2-mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50 µm. n.d. = not detected. Error bars depict SEM.
    Figure Legend Snippet: Rabbit ACE2-mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authentic virus (B) infection of Cos-7 cells transfected with ACE2 of various species. Infectivity was quantified by staining live virus with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2-mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50 µm. n.d. = not detected. Error bars depict SEM.

    Techniques Used: Infection, Transfection, Staining, Imaging

    Histological analysis of SARS-CoV-2 infected rabbits. Histopathological analysis of rabbits inoculated with 10 6 TCID 50 , sacrificed after four days. (A) Alveolar thickening and inflammatory infiltrates. Scale indicates 100 µm (B) Enlarged, syncytial cells in the alveolar lumina. Scale indicates 20 µm. (C) Lung pathology overview. Arrow indicates thickening and asterisk bronchus-associated lymphoid tissue (BALT). Scale indicates 200 µm. (D) Eosinophilic infiltrates in the nose. Scale indicates 40 µm.
    Figure Legend Snippet: Histological analysis of SARS-CoV-2 infected rabbits. Histopathological analysis of rabbits inoculated with 10 6 TCID 50 , sacrificed after four days. (A) Alveolar thickening and inflammatory infiltrates. Scale indicates 100 µm (B) Enlarged, syncytial cells in the alveolar lumina. Scale indicates 20 µm. (C) Lung pathology overview. Arrow indicates thickening and asterisk bronchus-associated lymphoid tissue (BALT). Scale indicates 200 µm. (D) Eosinophilic infiltrates in the nose. Scale indicates 40 µm.

    Techniques Used: Infection

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    Staining:

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    Sino Biological sars cov sars cov 2 nucleocapsid antibody mouse mab
    Counter screening of confirmed HCoV 229E hits with <t>SARS-CoV-2.</t> Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.
    Sars Cov Sars Cov 2 Nucleocapsid Antibody Mouse Mab, supplied by Sino Biological, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Sino Biological sars cov 2 nucleoprotein
    <t>SARS-CoV-2</t> nucleoprotein was present in several tissues of COVID-19-negative donors. ( a ) Representative images of donor tissue specimens analyzed by Western blot. Labels in the images show the location of tissue sampling in each organ/tissue. ( b ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from tissues of 8 different deceased donor PVW specimens, RV specimens from healthy donors (collected prior to COVID-19 pandemic), and LV specimen from COVID-19-positive deceased patient. GAPDH was used as a loading control protein. ( c , d ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from different cardiac and valve tissues of 2 deceased donors (#20-22 and #20-24) positive for SARS-CoV-2 serological assay. RV specimen from a healthy donor (collected prior to COVID-19 pandemic) and LV specimen from a COVID-19-positive deceased patient were used as controls. GAPDH was used as the loading control protein. PVW: pulmonary vein wall; RA: right atrium; IAS: interatrial septum; LA: left atrium; RV: right ventricle; IVS: interventricular septum; LV: left ventricle; AW: aortic wall; AVL: aortic valve leaflet.
    Sars Cov 2 Nucleoprotein, supplied by Sino Biological, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Counter screening of confirmed HCoV 229E hits with SARS-CoV-2. Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.

    Journal: bioRxiv

    Article Title: Repurposing screen highlights broad-spectrum coronavirus antivirals and their host targets

    doi: 10.1101/2021.07.14.452343

    Figure Lengend Snippet: Counter screening of confirmed HCoV 229E hits with SARS-CoV-2. Calu-3 cells were infected with SARS-CoV-2 (strain SARS-CoV-2/München-1.2/2020/984, p4) in the presence or absence of indicated compounds (final concentration 5 µM). (A) Infection efficiency was analyzed 48h post inoculation by quantification of viral genome equivalents in the culture fluid of cells. Culture fluid of cells inoculated with heat-inactivated SARS-CoV-2 served as background control (light blue circles). Compounds, which reduced virus load more than two standard deviations below the mean value of the DMSO control, were recognized as candidate antivirals. (B) Immunofluorescene analysis of infected cells. Cellular DNA was stained with DAPI (blue), and SARS-CoV-2 nucleoprotein with a monoclonal antibody (green). (C) Imaging-based quantification of total cell numbers and infected cell numbers. Images were automatically quantified as outlined in the methods section. The relative number of infected cells (N protein expressing cells) is correlated against the number of total cells (scale bar 125 µm). Means of four images per well of one experiment are given. DMSO and heat inactivated virus controls are depicted with light green and light blue circles, respectively. Data from Remdesivir-treated calls are depicted with red circles. Validated broad-spectrum coronavirus antivirals meeting our inclusion criteria are shown as with dark blue circles. Inclusion criterion was reduction of virus infected cell numbers by more than 50% and residual total cell number of more than 75% of DMSO solvent control. (D) Venn diagram highlighting the total number of qRT-PCR-based broad-spectrum candidates and confirmed hits based on the immunofluorescence analysis. Annotated targets of molecules with broad activity against both HCoV 229E and SARS-CoV-2 are given at the right. n.d., no data.

    Article Snippet: Immunofluorescence staining was conducted using the SARS-CoV-2 nucleoprotein mouse monoclonal antibody (1:500; Sino Biological #40143-MM05), Alexa-Fluor Plus 488 anti-mouse antibody (1:1,000; Invitrogen #A32723) and DAPI (1:10,000; Invitrogen #D21490).

    Techniques: Infection, Concentration Assay, Staining, Imaging, Expressing, Quantitative RT-PCR, Immunofluorescence, Activity Assay

    Results of ReFRAME library screening against the alphacoronavirus HCoV 229E. (A) Results of the dual luciferase-based HCoV 229E/R-Luc screening of the ReFRAME library in Huh-7.5/F-Luc cells. Renilla luciferase (R-Luc) activity is proportional to virus infection, whereas firefly luciferase (F-Luc) activity corresponds to cell numbers and viability. R-Luc and F-Luc values detected in DMSO-treated HCoV 229E infected cells were set to 100% and corrected for background values in case of R-Luc. (B) Stage of development of confirmed HCoV 229E hit compounds and information about previous screens by others. (C) Fifty-nine distinct host targets are annotated to these hits. The number before the dash indicates the number of antiviral hits with this target annotation, the number after the dash is equivalent to the total number of molecules within this target category in the ReFRAME library. (D) Overlap between this present ReFRAME library screening with HCoV 229E confirmed hits of the previous SARS-CoV-2 screening by Riva at al. ( Riva et al., 2020 ). Numbers indicate the IC50 ranks of the respective studies. (E) Dose-dependent antiviral activity of top ranked confirmed hits of Riva et al. against HCoV 229E. Dose-dependent antiviral activity of CDK inhibitors (F) and three inflammatory modulators (G) against HCoV 229E. Numbers before compound name indicate IC50-rank in the HCoV 229E screening. One experiment with means of two independent luciferase measurements normalized to DMSO control is given.

    Journal: bioRxiv

    Article Title: Repurposing screen highlights broad-spectrum coronavirus antivirals and their host targets

    doi: 10.1101/2021.07.14.452343

    Figure Lengend Snippet: Results of ReFRAME library screening against the alphacoronavirus HCoV 229E. (A) Results of the dual luciferase-based HCoV 229E/R-Luc screening of the ReFRAME library in Huh-7.5/F-Luc cells. Renilla luciferase (R-Luc) activity is proportional to virus infection, whereas firefly luciferase (F-Luc) activity corresponds to cell numbers and viability. R-Luc and F-Luc values detected in DMSO-treated HCoV 229E infected cells were set to 100% and corrected for background values in case of R-Luc. (B) Stage of development of confirmed HCoV 229E hit compounds and information about previous screens by others. (C) Fifty-nine distinct host targets are annotated to these hits. The number before the dash indicates the number of antiviral hits with this target annotation, the number after the dash is equivalent to the total number of molecules within this target category in the ReFRAME library. (D) Overlap between this present ReFRAME library screening with HCoV 229E confirmed hits of the previous SARS-CoV-2 screening by Riva at al. ( Riva et al., 2020 ). Numbers indicate the IC50 ranks of the respective studies. (E) Dose-dependent antiviral activity of top ranked confirmed hits of Riva et al. against HCoV 229E. Dose-dependent antiviral activity of CDK inhibitors (F) and three inflammatory modulators (G) against HCoV 229E. Numbers before compound name indicate IC50-rank in the HCoV 229E screening. One experiment with means of two independent luciferase measurements normalized to DMSO control is given.

    Article Snippet: Immunofluorescence staining was conducted using the SARS-CoV-2 nucleoprotein mouse monoclonal antibody (1:500; Sino Biological #40143-MM05), Alexa-Fluor Plus 488 anti-mouse antibody (1:1,000; Invitrogen #A32723) and DAPI (1:10,000; Invitrogen #D21490).

    Techniques: Library Screening, Luciferase, Activity Assay, Infection

    Validation of candidates with broad-spectrum coronavirus activity in well differentiated primary human lung cells. Primary cells from three independent donors were differentiated and grown as pseudostratified epithelium in air liquid interface culture. Cells were pre-treated for 2h with final concentrations of 1 µM (bright blue) or 5 µM (dark blue) of compounds from the basolateral side prior to inoculation with SARS-CoV-2 in the presence of compounds (apical and basal treatment). Infection efficiency was determined 72h post inoculation by qRT-PCR of cell extracts (A) and culture fluids (B). (C) Cell viability was assessed by quantification of GAPDH mRNA. Ct values > 35 were excluded from the analysis. Mean values (bar) and individual values (points) of three donors are given. Analysis was performed in two independent experiments (as indicated by the separation). In both experiments, primary cells from three different donors were used. Virus stocks differed in titer between these experiments (see Material and Methods).

    Journal: bioRxiv

    Article Title: Repurposing screen highlights broad-spectrum coronavirus antivirals and their host targets

    doi: 10.1101/2021.07.14.452343

    Figure Lengend Snippet: Validation of candidates with broad-spectrum coronavirus activity in well differentiated primary human lung cells. Primary cells from three independent donors were differentiated and grown as pseudostratified epithelium in air liquid interface culture. Cells were pre-treated for 2h with final concentrations of 1 µM (bright blue) or 5 µM (dark blue) of compounds from the basolateral side prior to inoculation with SARS-CoV-2 in the presence of compounds (apical and basal treatment). Infection efficiency was determined 72h post inoculation by qRT-PCR of cell extracts (A) and culture fluids (B). (C) Cell viability was assessed by quantification of GAPDH mRNA. Ct values > 35 were excluded from the analysis. Mean values (bar) and individual values (points) of three donors are given. Analysis was performed in two independent experiments (as indicated by the separation). In both experiments, primary cells from three different donors were used. Virus stocks differed in titer between these experiments (see Material and Methods).

    Article Snippet: Immunofluorescence staining was conducted using the SARS-CoV-2 nucleoprotein mouse monoclonal antibody (1:500; Sino Biological #40143-MM05), Alexa-Fluor Plus 488 anti-mouse antibody (1:1,000; Invitrogen #A32723) and DAPI (1:10,000; Invitrogen #D21490).

    Techniques: Activity Assay, Infection, Quantitative RT-PCR

    Rabbit ACE2 mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authen tic virus (B) infection of Cos-7 cells expressing ACE2 of various species. Infectivity was quantified by staining live virus cells with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2 mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50μm.

    Journal: bioRxiv

    Article Title: Susceptibility of rabbits to SARS-CoV-2

    doi: 10.1101/2020.08.27.263988

    Figure Lengend Snippet: Rabbit ACE2 mediated SARS-CoV-2 infection. SARS-CoV-2 pseudovirus (A) and authen tic virus (B) infection of Cos-7 cells expressing ACE2 of various species. Infectivity was quantified by staining live virus cells with anti-SARS-CoV nucleocapsid and scanning live virus and pseudovirus infected cells. (C) Confocal imaging of ACE2 mediated live virus infection; cells were stained using anti-human ACE2 in green, anti-SARS-CoV nucleocapsid in red and TO-PRO3 in blue to stain nuclei. Scale indicates 50μm.

    Article Snippet: After incubation, cells were formalin fixed, permeabilized with 70% ethanol and stained with 1:1000 mouse anti-SARS nucleoprotein (Sino Biological) and 1:1000 rabbit anti-human ACE2 (Abcam), followed by 1:1000 goat anti-rabbit Alexa-Fluor 594, 1:1000 goat anti-mouse Alexa-Fluor 488, and 1:1000 TO-PRO3 (Thermo Fisher) to stain nuclei.

    Techniques: Infection, Expressing, Staining, Imaging

    SARS-CoV-2 nucleoprotein was present in several tissues of COVID-19-negative donors. ( a ) Representative images of donor tissue specimens analyzed by Western blot. Labels in the images show the location of tissue sampling in each organ/tissue. ( b ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from tissues of 8 different deceased donor PVW specimens, RV specimens from healthy donors (collected prior to COVID-19 pandemic), and LV specimen from COVID-19-positive deceased patient. GAPDH was used as a loading control protein. ( c , d ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from different cardiac and valve tissues of 2 deceased donors (#20-22 and #20-24) positive for SARS-CoV-2 serological assay. RV specimen from a healthy donor (collected prior to COVID-19 pandemic) and LV specimen from a COVID-19-positive deceased patient were used as controls. GAPDH was used as the loading control protein. PVW: pulmonary vein wall; RA: right atrium; IAS: interatrial septum; LA: left atrium; RV: right ventricle; IVS: interventricular septum; LV: left ventricle; AW: aortic wall; AVL: aortic valve leaflet.

    Journal: Diagnostics

    Article Title: Presence of SARS-CoV-2 Nucleoprotein in Cardiac Tissues of Donors with Negative COVID-19 Molecular Tests

    doi: 10.3390/diagnostics11040731

    Figure Lengend Snippet: SARS-CoV-2 nucleoprotein was present in several tissues of COVID-19-negative donors. ( a ) Representative images of donor tissue specimens analyzed by Western blot. Labels in the images show the location of tissue sampling in each organ/tissue. ( b ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from tissues of 8 different deceased donor PVW specimens, RV specimens from healthy donors (collected prior to COVID-19 pandemic), and LV specimen from COVID-19-positive deceased patient. GAPDH was used as a loading control protein. ( c , d ) Western blot analysis for SARS-CoV-2 nucleoprotein on total protein extracts from different cardiac and valve tissues of 2 deceased donors (#20-22 and #20-24) positive for SARS-CoV-2 serological assay. RV specimen from a healthy donor (collected prior to COVID-19 pandemic) and LV specimen from a COVID-19-positive deceased patient were used as controls. GAPDH was used as the loading control protein. PVW: pulmonary vein wall; RA: right atrium; IAS: interatrial septum; LA: left atrium; RV: right ventricle; IVS: interventricular septum; LV: left ventricle; AW: aortic wall; AVL: aortic valve leaflet.

    Article Snippet: The in-depth analysis in cardiac specimens of donors #20-22 and #20-24 further highlighted the presence of SARS-CoV-2 nucleoprotein in other heart regions, especially in the right atrium and right ventricle (RA and RV).

    Techniques: Western Blot, Sampling, Serologic Assay

    Biodistribution of Sad23L-nCoV-S and Ad49L-nCoV-S vaccines in inoculated animals. ( A ) Serum NAb titers to Sad23L and Ad49L vectors were measured in macaques immunized by prime-boost inoculation with two vaccines at 4 week interval, or ( B ) in C57BL/6 and BALB/c mice 4 weeks post prime only or prime-boost vaccination with two vaccines or vectorial controls. ( C ) Nested-PCR amplification of Sad23L or Ad49L-hexon gene (500bp) in tissues of C57BL/6 mice 4 weeks after inoculation by prime only or prime-boost immunization with Sad23L-nCoV-S and Ad49L-nCoV-S vaccines. ( D ) Expression of S protein in splenocytes and hepatocytes of tissue frozen sections from vaccine immunized or control C57BL/6 mice by immunofluorescence staining with a human monoclonal antibody to SARS-CoV-2 S and DAPI.

    Journal: bioRxiv

    Article Title: Prime-boost vaccination of mice and Rhesus macaques with two novel adenovirus vectored COVID-19 vaccine candidates

    doi: 10.1101/2020.09.28.311480

    Figure Lengend Snippet: Biodistribution of Sad23L-nCoV-S and Ad49L-nCoV-S vaccines in inoculated animals. ( A ) Serum NAb titers to Sad23L and Ad49L vectors were measured in macaques immunized by prime-boost inoculation with two vaccines at 4 week interval, or ( B ) in C57BL/6 and BALB/c mice 4 weeks post prime only or prime-boost vaccination with two vaccines or vectorial controls. ( C ) Nested-PCR amplification of Sad23L or Ad49L-hexon gene (500bp) in tissues of C57BL/6 mice 4 weeks after inoculation by prime only or prime-boost immunization with Sad23L-nCoV-S and Ad49L-nCoV-S vaccines. ( D ) Expression of S protein in splenocytes and hepatocytes of tissue frozen sections from vaccine immunized or control C57BL/6 mice by immunofluorescence staining with a human monoclonal antibody to SARS-CoV-2 S and DAPI.

    Article Snippet: The expression of SARS-CoV-2 S protein was analyzed by Western blotting with rabbit polyclonal antibody to SARS-CoV-2 RBD (Sino Biological, China) and heat-inactivated human serum samples from Chinese COVID-19 infected patients.

    Techniques: Mouse Assay, Nested PCR, Amplification, Expressing, Immunofluorescence, Staining

    Examination of SARS-CoV-2 S protein in the tissues of Sad23L-nCoV-S and Ad49L-nCoV-S immunized mice. Spleen, liver, Lung and muscle tissues of immunized C57BL/6 mice were examined by immunofluorescence staining with a human monoclonal antibody to SARS-CoV-2 S and DAPI.

    Journal: bioRxiv

    Article Title: Prime-boost vaccination of mice and Rhesus macaques with two novel adenovirus vectored COVID-19 vaccine candidates

    doi: 10.1101/2020.09.28.311480

    Figure Lengend Snippet: Examination of SARS-CoV-2 S protein in the tissues of Sad23L-nCoV-S and Ad49L-nCoV-S immunized mice. Spleen, liver, Lung and muscle tissues of immunized C57BL/6 mice were examined by immunofluorescence staining with a human monoclonal antibody to SARS-CoV-2 S and DAPI.

    Article Snippet: The expression of SARS-CoV-2 S protein was analyzed by Western blotting with rabbit polyclonal antibody to SARS-CoV-2 RBD (Sino Biological, China) and heat-inactivated human serum samples from Chinese COVID-19 infected patients.

    Techniques: Mouse Assay, Immunofluorescence, Staining