Coiled-coil heterodimers with increased stability for cellular regulation and sensing SARS-CoV-2 spike protein-mediated cell fusion

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1. Version published to 10.1038/s41598-021-88315-3

   Apr 28, 2021
2. 

   ScreenIT

   Dec 11, 2020

   SciScore for 10.1101/2020.12.10.419440: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   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
   The primary antibodies were mouse anti-Myc (Cell signaling technologies; diluted 1:2,000), rabbit anti-HA (Sigma H6908; diluted 1:2,000), and mouse anti-β-actin (Cell Signaling 3700; diluted 1:2,000).	anti-Myc suggested: None anti-HA suggested: (Sigma-Aldrich Cat# H6908, RRID:AB_260070) anti-β-actin suggested: None
   The secondary antibodies were HRP-conjugated goat anti-rabbit IgG (Abcam ab6721; diluted 1:3,000) and HRP-conjugated goat anti-mouse IgG (Santa Cruz, …

   SciScore for 10.1101/2020.12.10.419440: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   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
   The primary antibodies were mouse anti-Myc (Cell signaling technologies; diluted 1:2,000), rabbit anti-HA (Sigma H6908; diluted 1:2,000), and mouse anti-β-actin (Cell Signaling 3700; diluted 1:2,000).	anti-Myc suggested: None anti-HA suggested: (Sigma-Aldrich Cat# H6908, RRID:AB_260070) anti-β-actin suggested: None
   The secondary antibodies were HRP-conjugated goat anti-rabbit IgG (Abcam ab6721; diluted 1:3,000) and HRP-conjugated goat anti-mouse IgG (Santa Cruz, sc2005; diluted 1:3,000).	anti-rabbit IgG suggested: (Abcam Cat# ab6721, RRID:AB_955447) anti-mouse IgG suggested: (Santa Cruz Biotechnology Cat# sc-2005, RRID:AB_631736)
   Experimental Models: Cell Lines
   Sentences	Resources
   At 70% confluency, the donor HEK293T cells were transfected with plasmids expressing S protein, syncytia reporter split fluorescent protein GFP1-10 (GFP1-10:N7, GFP1-10:P7 or GFP1-10) and fluorescent protein mCherryNLS, and the acceptor cells were transfected with plasmids expressing the ACE2 receptor, syncytia reporter split protein GFP11 (N8:GFP11, 3×(N8:GFP11), P8:GFP11), and fluorescent protein BFP using PEI transfection reagent.	HEK293T suggested: None
   Software and Algorithms
   Sentences	Resources
   The exact peptide concentrations were determined based on absorbance at 280 nm, and the extinction coefficients were calculated by the ProtParam web tool (https://web.expasy.org/cgi-bin/protparam/protparam).	ProtParam suggested: (ProtParam Tool, RRID:SCR_018087)
   Data analysis was carried out with Astra 7.0 software (Wyatt, CA) utilizing the theoretical extinction coefficient calculated by ProtParam.	Astra suggested: (ASTRA, RRID:SCR_016255)
   The integration of heat pulses and model-fitting were performed with Origin 7.0 using a single-site binding model.	Origin suggested: (Origin, RRID:SCR_014212)
   Data were analyzed using FlowJo (TreeStar, Ashland, OR, USA) and SpectroFlo (Cytek) software.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   The images were processed with LAS AF software (Leica Microsystems) and ImageJ software (National Institute of Mental Health, Bethesda, Maryland,	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Software and statistics: Graphs were prepared with Origin 8.1 (http://www.originlab.com/), and GraphPad Prism 5 (http://www.graphpad.com/) was used for statistical purposes.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   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).

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   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.

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   Results from TrialIdentifier: No clinical trial numbers were referenced.

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   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).

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

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   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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3. 

   Version published to 10.1101/2020.12.10.419440v1 on bioRxiv

   Dec 10, 2020