Receptome profiling identifies KREMEN1 and ASGR1 as alternative functional receptors of SARS-CoV-2

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Abstract

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  1. SciScore for 10.1101/2020.09.09.287508: (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
    Supernatant was discarded after centrifugation and washed once with PBS/2%FBS, the cells were then labelled with Anti-hFc-APC (Jackson Lab) antibody for 20min, and washed once with PBS/2%FBS.
    Anti-hFc-APC (Jackson Lab)
    suggested: None
    Beads were washed three times with the RIPA buffer, and the samples were prepared for western blot with anti-hFc or anti-FLAG antibodies.
    anti-hFc
    suggested: None
    anti-FLAG
    suggested: None
    SARS-CoV-2 replication was examined by immuno-fluorescence and flow cytometry with anti SARS-CoV-2 N protein antibody.
    anti SARS-CoV-2 N protein
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Cell culture and transfection: Vero E6 cells and HEK293T cells were cultured in DMEM supplemented with 10% FBS at 37°C in 5% CO2 and the normal level of O2.
    HEK293T
    suggested: None
    HEK293e cells were cultured in serum-free FreeStyle 293 Medium (Invitrogen) with 120 rpm rotation at 37°C in 5% CO2 and the normal level of O2.
    HEK293e
    suggested: None
    ACE2 knockout 293T stable cell line: ACE2 small guide RNA was constructed into pSLQ1651 (Addgene #51024) (44) with a targeting sequence of CTTGGCCTGTTCCTCAATGGTGG.
    293T
    suggested: None
    ACE2 KO 293T stable cell line were obtained by single cell dilution.
    ACE2 KO 293T
    suggested: None
    Authentic SARS-CoV-2 generation and infection: SARS-CoV-2/MT020880.1 were expanded in Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    The flow data were analyzed with FlowJo software.
    FlowJo
    suggested: (FlowJo, RRID:SCR_008520)
    Data analysis and statistics: Gene Ontology Enrichment Analysis was performed by R bioconductor.
    bioconductor
    suggested: (Bioconductor, RRID:SCR_006442)

    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 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.


    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.