A Replication-Competent Vesicular Stomatitis Virus for Studies of SARS-CoV-2 Spike-Mediated Cell Entry and Its Inhibition
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SciScore for 10.1101/2020.05.20.105247: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: After obtaining informed consent, serum was obtained by venipuncture (BD Vacutainer, serum), centrifuged, aliquoted and stored at -80°C prior to use.
IRB: Protocol approval was obtained by the Institutional Review Board (IRB) of the Albert Einstein College of Medicine.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of hACE2 in BHK21 transfected cells: To stain for surface-expressed hACE2, BHK21-hACE2 or the control BHK21 cells seeded onto fibronectin-coated glass coverslips were incubated with 0.4 … SciScore for 10.1101/2020.05.20.105247: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: After obtaining informed consent, serum was obtained by venipuncture (BD Vacutainer, serum), centrifuged, aliquoted and stored at -80°C prior to use.
IRB: Protocol approval was obtained by the Institutional Review Board (IRB) of the Albert Einstein College of Medicine.Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Detection of hACE2 in BHK21 transfected cells: To stain for surface-expressed hACE2, BHK21-hACE2 or the control BHK21 cells seeded onto fibronectin-coated glass coverslips were incubated with 0.4 μg/mL of hACE2-specific goat antibody (#AF933, R&D systems) at 4°C in media containing 25 mM HEPES. antibodysuggested: (R and D Systems Cat# AF933, RRID:AB_355722)Anti-hACE2 antibody blocking assay: Huh7.5.1 cells were seeded into a 384-well plate. Anti-hACE2suggested: NoneNex day, goat anti-human ACE2 antibody (#AF933, R&D Systems) was serially diluted and applied to Huh 7.5.1 cells. anti-human ACE2suggested: NoneCells were blocked for ∼2 h, and then immunostained with SARS-1 nucleocapsid protein-specific antibody (SinoBiologic; 40143-V08B) and Alexa Fluor-488 labeled secondary antibody. SARS-1 nucleocapsid protein-specificsuggested: NoneExperimental Models: Cell Lines Sentences Resources Vero-E6 cells were grown in MEM supplemented in 10% FBS and Gentimicine. Vero-E6suggested: NoneRetroviruses were produced by transfecting 293FT cells with the hACE2 and TMPRSS2 expressing pBabe-puro plasmids along with those expressing the Moloney murine leukemia virus (MMLV) gag-pol and VSV G proteins. 293FTsuggested: ATCC Cat# PTA-5077, RRID:CVCL_6911)Detection of hACE2 in BHK21 transfected cells: To stain for surface-expressed hACE2, BHK21-hACE2 or the control BHK21 cells seeded onto fibronectin-coated glass coverslips were incubated with 0.4 μg/mL of hACE2-specific goat antibody (#AF933, R&D systems) at 4°C in media containing 25 mM HEPES. BHK21suggested: NoneSerum:virus mixtures were then added in duplicate to 384-well plates (Corning) containing Huh7.5.1 cells or 96-well plates (Corning) containing Vero cells. Huh7.5.1suggested: RRID:CVCL_E049)Verosuggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)SARS-CoV-2 stock preparation: Vero-76 cells were inoculated with SARS-CoV-2 (GenBank MT020880.1) at an MOI of 0.01 and incubated at 37°C with 5% CO2 and 80% humidity. Vero-76suggested: NoneSoftware and Algorithms Sentences Resources Viral infectivity was measured by automated enumeration of GFP-positive cells from captured images using a Cytation5 automated fluorescence microscope (BioTek) and analyzed using the Gen5 data analysis software (BioTek). Gen5suggested: (Gen5, RRID:SCR_017317)The half-maximal inhibitory concentration (IC50) of the mAbs was calculated using a nonlinear regression analysis with GraphPad Prism software. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)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: Please consider improving the rainbow (“jet”) colormap(s) used on page 17. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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.
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