SARS-CoV-2 invades host cells via a novel route: CD147-spike protein

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   ScreenIT

   Jun 5, 2020

   SciScore for 10.1101/2020.03.14.988345: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Ethics statement: This study was approved by the ethics committee of First Affiliated Hospital of Fourth Military Medical University (KY20202005-1).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	Authentication: Cell culture: The cell lines (Vero E6 and 293T) have been tested and authenticated using Short Tandem Repeat DNA profiling by Beijing Microread Genetics Co., Ltd (Beijing, China) and were cultured at 37°C under 5% CO2 in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin and 2% L-glutamine.

   Table 2: Resources

   Antibodies
   S…

   SciScore for 10.1101/2020.03.14.988345: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: Ethics statement: This study was approved by the ethics committee of First Affiliated Hospital of Fourth Military Medical University (KY20202005-1).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	Authentication: Cell culture: The cell lines (Vero E6 and 293T) have been tested and authenticated using Short Tandem Repeat DNA profiling by Beijing Microread Genetics Co., Ltd (Beijing, China) and were cultured at 37°C under 5% CO2 in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin and 2% L-glutamine.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Anti-CD147 antibody (HAb18, produced by our laboratory) and anti-SARS-CoV-2 Spike antibody (40150-R007, Sino Biological, China) were used for antibody immobilization for Co-IP.	Anti-CD147 suggested: None
   After blocking with 5% non-fat milk for 1 hour, the membrane was incubated with the corresponding primary antibodies (anti-CD147, HAb18, produced by our laboratory, dilution 1:2000; anti-SARS-CoV-2 Spike antibody, 40150-R007, Sino Biological, China, dilution 1:2000) at 4°C overnight.	HAb18 suggested: None
   The images were developed after incubation with the secondary antibodies (goat anti-mouse IgG(H+L) antibody, 31430, Thermo Fisher Scientific, MA, USA, dilution	anti-mouse IgG(H+L suggested: None
   n 1:5000; goat anti-rabbit IgG(H+L) antibody; 31460, Thermo Fisher Scientific, MA, USA, dilution 1:5000) at room temperature for 1 hour.	anti-rabbit IgG(H+L) suggested: None
   After washing with PBST, the samples were incubated with anti-CD147 antibody (HAb18, produced by our laboratory) for 1 hour; and then incubated with horseradish peroxidase (HRP)-labeled goat anti-mouse antibody for 1 hour.	anti-mouse suggested: None
   Subsequently, the samples were incubated with anti-SARS-CoV-2 Spike antibody (40150-R007, Sino Biological, China) and then detected with HRP-labeled goat anti-rabbit antibody for 1 hour at 37°C, the following steps were similar to those described earlier.	anti-SARS-CoV-2 suggested: None anti-rabbit suggested: None
   Having been blocked with goat serum for 30min, the sections was incubated with corresponding primary antibodies (anti-CD147, HAb18, produced by our laboratory, dilution 1:200; anti-SARS-CoV-2 Spike antibody, 40150-R007, Sino Biological, China, dilution 1:400) for 16 hours at 4°C overnight.	anti-CD147, HAb18 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cell culture: The cell lines (Vero E6 and 293T) have been tested and authenticated using Short Tandem Repeat DNA profiling by Beijing Microread Genetics Co., Ltd (Beijing, China) and were cultured at 37°C under 5% CO2 in DMEM or RPMI 1640 medium supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin and 2% L-glutamine.	293T suggested: None
   Immuno-electron microscope: Vero E6 cells infected with SARS-CoV-2 were harvested and treated with fixative at 4°C.	Vero E6 suggested: RRID:CVCL_XD71)
   Software and Algorithms
   Sentences	Resources
   The results were analyzed by BIAevaluation software to determine the affinity constant.	BIAevaluation suggested: (BIAevaluation Software, RRID:SCR_015936)
   Statistical Analysis: All the statistical analyses were performed using GraphPad prism 5.0.	GraphPad 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 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:

   • No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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2. 

   Version published to 10.1101/2020.03.14.988345 on bioRxiv

   Mar 14, 2020