Spike Protein of SARS-CoV-2 Activates Macrophages and Contributes to Induction of Acute Lung Inflammations in Mice

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   ScreenIT

   Dec 8, 2020

   SciScore for 10.1101/2020.12.07.414706: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Intravenous viral administration in vivo: The animal protocol was approved by the University of South Carolina IACUC committee.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Male wild type C57BL/6J mice (5-6 weeks old) were intravenously administered 100 μl of Spp or VSV-G lentivirus (8×108 of viral particles) via tail venous or retro-orbital injection.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Anti-Spike S1 subunit, anti-low density lipoprotein receptor (LDLR), anti-mannose receptor C-type 1 (MRC1), anti-CD68 and anti-human immunodeficiency viruses (HIV)-1 p24 …

   SciScore for 10.1101/2020.12.07.414706: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: Intravenous viral administration in vivo: The animal protocol was approved by the University of South Carolina IACUC committee.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Male wild type C57BL/6J mice (5-6 weeks old) were intravenously administered 100 μl of Spp or VSV-G lentivirus (8×108 of viral particles) via tail venous or retro-orbital injection.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Anti-Spike S1 subunit, anti-low density lipoprotein receptor (LDLR), anti-mannose receptor C-type 1 (MRC1), anti-CD68 and anti-human immunodeficiency viruses (HIV)-1 p24 antibodies were obtained from Novus Biologicals (Littleton, CO, USA).	Anti-Spike S1 subunit, anti-low density lipoprotein receptor (LDLR), anti-mannose receptor C-type 1 suggested: None MRC1 suggested: None anti-CD68 suggested: None anti-human immunodeficiency suggested: None HIV)-1 p24 suggested: None
   The anti-His tag antibodies were obtained from Proteintech (Rosemont, IL, USA) and Thermo Fisher (Waltham, MA, USA).	The anti-His tag antibodies suggested: None anti-His tag suggested: None
   Anti-Spike S1 subunit, anti-HIV-1 p24 antibody, or anti-His antibodies were used to detect the expression of viral proteins and followed by HRP-labeled secondary antibodies.	Anti-Spike S1 subunit, suggested: (Active Motif Cat# 91345, RRID:AB_2847847) anti-HIV-1 suggested: None anti-His suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   RAW264.7 cell culture, viral uptake and electroporation: Macrophage-like RAW264.7 (RAW) cells (ATCC® TIB-71™) were cultured in DMEM (10% FBS) medium.	RAW264.7 suggested: ATCC Cat# TIB-71, RRID:CVCL_0493)
   In a parallel experiment, RAW cells (5×106) were electroporated with pcDNA3.1, EGFP-N2 or pcDNA-Spike plasmids (10 μg) using the following parameters: 2 mm gap cuvette, 250 ul sample volume and 120V (BTX Harvard Bioscience, Inc., Holliston, MA, USA).	RAW suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Wild type C57BL/6J mice were purchased from the Jackson Laboratory (Bar Harbor, ME, USA).	C57BL/6J suggested: RRID:IMSR_JAX:000664)
   Software and Algorithms
   Sentences	Resources
   Images were acquired using ImageXpress Pico System (Molecular Device, San Jose, CA, USA) or confocal microscopy system (Carl Zeiss AG, Oberkochen, Germany).	ImageXpress Pico System suggested: None

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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 did not find any issues relating to the usage of bar graphs.

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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.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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   Read the original source
2. 

   Version published to 10.1101/2020.12.07.414706v1 on bioRxiv

   Dec 7, 2020