Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) membrane (M) protein inhibits type I and III interferon production by targeting RIG-I/MDA-5 signaling

  1. Yi Zheng
  2. Meng-Wei Zhuang
  3. Lulu Han
  4. Jing Zhang
  5. Mei-Ling Nan
  6. Peng Zhan
  7. Dongwei Kang
  8. Xinyong Liu
  9. Chengjiang Gao
  10. Pei-Hui Wang

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Abstract

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has quickly spread worldwide and has affected more than 10 million individuals. A typical feature of COVID-19 is the suppression of type I and III interferon (IFN)-mediated antiviral immunity. However, the molecular mechanism by which SARS-CoV-2 evades antiviral immunity remains elusive. Here, we reported that the SARS-CoV-2 membrane (M) protein inhibits the production of type I and III IFNs induced by the cytosolic dsRNA-sensing pathway mediated by RIG-I/MDA-5–MAVS signaling. In addition, the SARS-CoV-2 M protein suppresses type I and III IFN induction stimulated by SeV infection or poly (I:C) transfection. Mechanistically, the SARS-CoV-2 M protein interacts with RIG-I, MAVS, and TBK1, thus preventing the formation of the multiprotein complex containing RIG-I, MAVS, TRAF3, and TBK1 and subsequently impeding the phosphorylation, nuclear translocation, and activation of IRF3. Consequently, ectopic expression of the SARS-CoV-2 M protein facilitates the replication of vesicular stomatitis virus. Taken together, these results indicate that the SARS-CoV-2 M protein antagonizes type I and III IFN production by targeting RIG-I/MDA-5 signaling, which subsequently attenuates antiviral immunity and enhances viral replication. This study provides insight into the interpretation of SARS-CoV-2-induced antiviral immune suppression and illuminates the pathogenic mechanism of COVID-19.

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  1. Version published to 10.1038/s41392-020-00438-7
  2. ScreenIT

    SciScore for 10.1101/2020.07.26.222026: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    29 Antibodies and reagents: Rabbit anti-DYKDDDDK Tag (D6W5B), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2), and rabbit anti-TRAF3 were from Cell Signaling Technology (
    anti-DYKDDDDK
    suggested: (Cell Signaling Technology Cat# 14793, RRID:AB_2572291)
    anti-IRF3
    suggested: None
    anti-pIRF3 ( 4D46)
    suggested: None
    anti-TBK1
    suggested: (Cell Signaling Technology Cat# 14590, RRID:AB_2798527)
    anti-pTBK1
    suggested: None
    anti-TRAF3
    suggested: None
    Alexa Fluor 488 goat anti-rabbit IgG secondary antibody, Alexa Fluor 568 goat anti-mouse IgG secondary antibody, Alexa Fluor 488 goat anti-mouse IgG secondary antibody, and Alexa Fluor 568 goat anti-rabbit IgG secondary antibody were from Thermo Fisher Scientific (USA).
    Alexa Fluor 488 goat anti-rabbit IgG secondary antibody
    suggested: None
    anti-mouse IgG
    suggested: None
    anti-rabbit IgG
    suggested: None
    Coimmunoprecipitation and immunoblotting: For coimmunoprecipitation assays, HEK293T cells were collected 24 hours after transfection and lysed in lysis buffer [1.0% (v/v) NP-40, 50 mM Tris-HCl, pH 7.4, 50 mM EDTA, 0.15 M NaCl] supplemented with a protease inhibitor cocktail (Sigma, USA) and a phosphatase inhibitor cocktail (Sigma, USA) as described in our previous publications.30,31 After centrifugation for 10 min at 14,000 g, the supernatants were collected and incubated with the indicated antibodies, followed by the addition of protein A/G beads (Santa Cruz, USA)
    NP-40
    suggested: (Covance Cat# MMS-503P-100, RRID:AB_291448)
    Experimental Models: Cell Lines
    SentencesResources
    Cell culture and transfection: HEK293, HEK293T, HeLa, and Vero-E6 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Gibco, USA) with 10% heat-inactivated fetal bovine serum (FBS, Gibco, USA).
    Vero-E6
    suggested: None
    32,35 Briefly, approximately 0.5 x 105 HEK293T cells were seeded in 48-well plates and transfected 12 hours later with the luciferase reporter plasmid and the expression vector plasmids of RIG-I, RIG-IN, MDA-5, MAVS, TBK1, IKKε, IRF3-5D, TRIF, and STING, alone or together with the plasmid expressing the SARS-CoV-2 M protein, as indicated in the experiments.
    MDA-5
    suggested: None
    Viruses and infection: VSV-enhanced green fluorescent protein (eGFP) and SeV were used to infect HeLa, HEK293, or HEK293T cells as described in our previous publications.30–32 Briefly, before infection, prewarmed serum-free DMEM medium at 37°C was used to wash the target cells, after which the virus was diluted to the desired multiplicity of infection (MOI) in serum-free DMEM and incubated with the target cells for 1-2 hours.
    HEK293
    suggested: None
    HEK293T
    suggested: None
    Confocal immunofluorescence microscopy: Confocal immunofluorescence microscopy studies were performed as described in our previous publications.30,31 Briefly, HeLa cells were grown on 12-well slides one day before transfection with the indicated plasmids.
    HeLa
    suggested: None
    32 Briefly, Vero cells were seeded on 24-well plates.
    Vero
    suggested: None
    Software and Algorithms
    SentencesResources
    For statistical analysis, two-tailed unpaired Student’s t-tests were performed by GraphPad Prism 8.0 and Microsoft Excel.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.

    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

    Results from JetFighter: We did not find any issues relating to colormaps.

    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  3. ScreenIT

    SciScore for 10.1101/2020.07.26.222026: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.Randomizationnot detected.Blindingnot detected.Power Analysisnot detected.Sex as a biological variablenot detected.Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    29 Antibodies and reagents Rabbit anti-DYKDDDDK Tag (D6W5B), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2), and rabbit anti-TRAF3 were from Cell Signaling Technology (
    anti-DYKDDDDK
    suggested: (Cell Signaling Technology Cat# 14793, RRID:AB_2572291)
    anti-IRF3
    suggested: None
    anti-pIRF3 ( 4D46)
    suggested: None
    anti-TBK1
    suggested: (Cell Signaling Technology Cat# 14590, RRID:AB_2798527)
    anti-pTBK1
    suggested: None
    anti-TRAF3
    suggested: None
    Alexa Fluor 488 goat anti-rabbit IgG secondary antibody, Alexa Fluor 568 goat anti-mouse IgG secondary antibody, Alexa Fluor 488 goat anti-mouse 10 / 55 IgG secondary antibody, and Alexa Fluor 568 goat anti-rabbit IgG secondary antibody were from Thermo Fisher Scientific (USA).
    Alexa Fluor 488 goat anti-rabbit IgG secondary antibody
    suggested: None
    anti-mouse IgG
    suggested: None
    anti-mouse 10 / 55 IgG
    suggested: None
    anti-rabbit IgG
    suggested: None
    Coimmunoprecipitation and immunoblotting For coimmunoprecipitation assays, HEK293T cells were collected 24 hours after transfection and lysed in lysis buffer [1.0% (v/v) NP-40, 50 mM Tris-HCl, pH 7.4, 50 mM EDTA, 0.15 M NaCl] supplemented with a protease inhibitor cocktail (Sigma, USA) and a phosphatase inhibitor cocktail (Sigma, USA) as described in our previous publications.30,31 After centrifugation for 10 min at 14,000 g, the supernatants were collected and incubated with the indicated 13 / 55 antibodies, followed by the addition of protein A/G beads (Santa Cruz, USA)
    NP-40
    suggested: (Covance Cat# MMS-503P-100, RRID:AB_291448)
    The expression plasmids of the SARS-CoV-2 M protein and plasmids expressing RIG-I or MDA-5 were cotransfected into HEK293T cell, 24 hours later, MAVS antibodies were used to perform coimmunoprecipitation.
    MAVS
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    9 / 55 Materials and methods Cell culture and transfection HEK293, HEK293T, HeLa, and Vero-E6 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, USA) with 10% heat-inactivated fetal bovine serum (FBS, Gibco, USA).
    Vero-E6
    suggested: None
    32,35 Briefly, approximately 0.5 x 105 HEK293T cells were seeded in 48-well plates and transfected 12 hours later with the luciferase reporter plasmid and the expression vector plasmids of RIG-I, RIG-IN, MDA-5, MAVS, TBK1, IKK , IRF3-5D, TRIF, and STING, alone or together with the plasmid ε expressing the SARS-CoV-2 M protein, as indicated in the experiments.
    MDA-5
    suggested: None
    Viruses and infection VSV-enhanced green fluorescent protein (eGFP) and SeV were used to infect HeLa, HEK293, or HEK293T cells as described in our previous publications.30-32 Briefly, before infection, prewarmed serum-free DMEM medium at 37°C was used to wash the target cells, after which the virus was diluted to the desired multiplicity of infection (MOI) in serum-free DMEM and incubated with the target cells for 1-2 hours.
    HeLa
    suggested: None
    HEK293
    suggested: None
    32 Briefly, Vero cells were seeded on 24-well plates.
    Vero
    suggested: None
    In a ll cases,ava lue ofP<0.0 5was conside red tobestatisti callysi gnifican t.16/ 55R esultsT heSAR S-Co V-2Mprot ei ninhibitst ypeIandI IIIFNin ductio nbySeV andpol y(I: C)To exp lorewhethertheSA RS-Co V-2Mpr otein affec tstype IandI III FNpro duct ion,HEK2 93 Tcel ls e xpressingt heSARS-Co V-2M pr otei nwe reinfect edwi thSeVortran sfec tedwith adsRNAmi mic ,pol y(I:C ).
    HEK293T
    suggested: KCB Cat# KCB 200744YJ, RRID:CVCL_0063
    Software and Algorithms
    SentencesResources
    After removing the solid agarose-medium mix, the cells were stained with 0.05% crystal violet, and the plaques on the monolayer were then counted to calculate the virus titer. 15 / 55 Bioinformatics analysis The transmembrane motifs were predicted with the TMHMM server version 2.0 (http://www.cbs.dtu.dk/services/TMHMM/).
    http://www.cbs.dtu.dk/services/TMHMM/
    suggested: (TMHMM Server, RRID:SCR_014935)
    For statistical analysis, two-tailed unpaired Student's t-tests were performed by GraphPad Prism 8.0 and Microsoft Excel.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    Microsoft Excel
    suggested: (Microsoft Excel, RRID:SCR_016137)

    Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).

    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from Barzooka: We found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

    Results from JetFighter: We did not find any issues relating to colormaps.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  4. ScreenIT

    SciScore for 10.1101/2020.07.26.222026: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.Randomizationnot detected.Blindingnot detected.Power Analysisnot detected.Sex as a biological variablenot detected.Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    29 Antibodies and reagents Rabbit anti-DYKDDDDK Tag (D6W5B), rabbit anti-IRF3 (D83B9), rabbit anti-pIRF3 (4D46), rabbit anti-TBK1 (3031S), rabbit anti-pTBK1 (D52C2), and rabbit anti-TRAF3 were from Cell Signaling Technology (
    anti-DYKDDDDK
    suggested: (Cell Signaling Technology Cat# 14793, AB_2572291)
          <div style="margin-bottom:8px">
        <div><b>anti-IRF3</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-pIRF3 ( 4D46)</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-pTBK1</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-TRAF3</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Alexa Fluor 488 goat anti-rabbit IgG secondary antibody, Alexa Fluor 568 goat anti-mouse IgG secondary antibody, Alexa Fluor 488 goat anti-mouse 10 / 55 IgG secondary antibody, and Alexa Fluor 568 goat anti-rabbit IgG secondary antibody were from Thermo Fisher Scientific (USA).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Alexa Fluor 488 goat anti-rabbit IgG secondary antibody</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-mouse IgG</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-mouse 10 / 55 IgG</b></div>
        <div>suggested: None</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>anti-rabbit IgG</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Coimmunoprecipitation and immunoblotting For coimmunoprecipitation assays, HEK293T cells were collected 24 hours after transfection and lysed in lysis buffer [1.0% (v/v) NP-40, 50 mM Tris-HCl, pH 7.4, 50 mM EDTA, 0.15 M NaCl] supplemented with a protease inhibitor cocktail (Sigma, USA) and a phosphatase inhibitor cocktail (Sigma, USA) as described in our previous publications.30,31 After centrifugation for 10 min at 14,000 g, the supernatants were collected and incubated with the indicated 13 / 55 antibodies, followed by the addition of protein A/G beads (Santa Cruz, USA)</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>NP-40</b></div>
        <div>suggested: (Covance Cat# MMS-503P-100, <a href="https://scicrunch.org/resources/Any/search?q=AB_291448">AB_291448</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The expression plasmids of the SARS-CoV-2 M protein and plasmids expressing RIG-I or MDA-5 were cotransfected into HEK293T cell, 24 hours later, MAVS antibodies were used to perform coimmunoprecipitation.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>MAVS</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">When using the TBK1 antibody to perform endogenous coimmunoprecipitation, MAVS was detected in the TBK1 immunoprecipitates (Fig. 5c).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>TBK1</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Experimental Models: Cell Lines</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">12 The binding of the type I or III IFNs to their specific receptors, the type I IFN receptor (IFNAR) and the type III IFN receptor (IFNLR), respectively, triggers the activation of the receptor-associated Janus kinase 1 (JAK1)/tyrosine kinase 2 (TYK2), which stimulates the phosphorylation of STAT1 and STAT2.9,13 JAK2 also participates in type III IFN-induced STAT phosphorylation.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>STAT2.9,13 JAK2</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">This study reveals a previously undiscovered mechanism of SARS-CoV-2 in evading host antiviral immunity, which may partially explain the clinical features of impaired antiviral immunity in COVID-19 patients and provide insights into the viral pathogenicity and treatment. 9 / 55 Materials and methods Cell culture and transfection HEK293, HEK293T, HeLa, and Vero-E6 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, Gibco, USA) with 10% heat-inactivated fetal bovine serum (FBS, Gibco, USA).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero-E6</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">Viruses and infection VSV-enhanced green fluorescent protein (eGFP) and SeV were used to infect HeLa, HEK293, or HEK293T cells as described in our previous publications.30-32 Briefly, before infection, prewarmed serum-free DMEM medium at 37°C was used to wash the target cells, after which the virus was diluted to the desired multiplicity of infection (MOI) in serum-free DMEM and incubated with the target cells for 1-2 hours.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>HEK293</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">32 Briefly, Vero cells were seeded on 24-well plates.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>Vero</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">The results indicated that both SeV infection and poly (I:C) transfection strongly stimulated the expression of IFN- , IFN- 1, β λ ISG56, and CXCL10 in the control HEK293T cells (Fig. 1).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>HEK293T</b></div>
        <div>suggested: KCB Cat# KCB 200744YJ, <a href="https://scicrunch.org/resources/Any/search?q=CVCL_0063">CVCL_0063</a></div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">When the SARS-CoV-2 M protein was overexpressed, the binding between RIG-I and MAVS was reduced (Fig. 5a, lanes 2 compared to lane 3); however, in the same condition, the interaction between MDA-5 and MAVS was not affected (Fig 5b, lanes 2 compared to lane 3), indicating that the SARS-CoV-2 M protein impedes the complex formation of RIG-I and MAVS but has no effect on the interaction between MDA-5 and MAVS.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>MDA-5</b></div>
        <div>suggested: None</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">To address the effect of the SARS-CoV-2 M protein on virus-induced IRF3 phosphorylation, control HeLa cells and HeLa cells expressing the SARS-CoV-2 M protein were infected with SeV.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>HeLa</b></div>
        <div>suggested: CLS Cat# 300194/p772_HeLa, <a href="https://scicrunch.org/resources/Any/search?q=CVCL_0030">CVCL_0030</a></div>
      </div>
    </td></tr><tr><td style="min-width:100px;text-align:center; padding-top:4px;" colspan="2"><b>Software and Algorithms</b></td></tr><tr><td style="min-width:100px;text=align:center"><i>Sentences</i></td><td style="min-width:100px;text-align:center"><i>Resources</i></td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">After removing the solid agarose-medium mix, the cells were stained with 0.05% crystal violet, and the plaques on the monolayer were then counted to calculate the virus titer. 15 / 55 Bioinformatics analysis The transmembrane motifs were predicted with the TMHMM server version 2.0 (http://www.cbs.dtu.dk/services/TMHMM/).</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>http://www.cbs.dtu.dk/services/TMHMM/</b></div>
        <div>suggested: (TMHMM Server, <a href="https://scicrunch.org/resources/Any/search?q=SCR_014935">SCR_014935</a>)</div>
      </div>
    </td></tr><tr><td style="min-width:100px;vertical-align:top;border-bottom:1px solid lightgray">For statistical analysis, two-tailed unpaired Student's t-tests were performed by GraphPad Prism 8.0 and Microsoft Excel.</td><td style="min-width:100px;border-bottom:1px solid lightgray">
      <div style="margin-bottom:8px">
        <div><b>GraphPad Prism</b></div>
        <div>suggested: (GraphPad Prism, <a href="https://scicrunch.org/resources/Any/search?q=SCR_002798">SCR_002798</a>)</div>
      </div>
    
      <div style="margin-bottom:8px">
        <div><b>Microsoft Excel</b></div>
        <div>suggested: (Microsoft Excel, <a href="https://scicrunch.org/resources/Any/search?q=SCR_016137">SCR_016137</a>)</div>
      </div>
    </td></tr></table>

    Data from additional tools added to each annotation on a weekly basis.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore is not a substitute for expert review. SciScore checks for the presence and correctness of RRIDs (research resource identifiers) in the manuscript, and detects sentences that appear to be missing RRIDs. SciScore also checks to make sure that rigor criteria are addressed by authors. It does this by detecting sentences that discuss criteria such as blinding or power analysis. SciScore does not guarantee that the rigor criteria that it detects are appropriate for the particular study. Instead it assists authors, editors, and reviewers by drawing attention to sections of the manuscript that contain or should contain various rigor criteria and key resources. For details on the results shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2020.07.26.222026 on bioRxiv