Bromelain Inhibits SARS-CoV-2 Infection in VeroE6 Cells

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1. 

   ScreenIT

   Sep 21, 2020

   SciScore for 10.1101/2020.09.16.297366: (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
   Further, blocked with 5% skimmed milk powder at room temperature for 1 h, the membrane was incubated with the target primary antibody with desired dilution (anti-ACE-2, 1:1000	anti-ACE-2 suggested: None
   For loading, control membranes were probed with anti-β-actin and anti-GAPDH antibodies (1:5000, Cell signaling technology, MA, USA).	anti-β-actin suggested: None anti-GAPDH suggested: None
   The membrane was washed with 1X TBST (3 × 5 min) and then incubated with …

   SciScore for 10.1101/2020.09.16.297366: (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
   Further, blocked with 5% skimmed milk powder at room temperature for 1 h, the membrane was incubated with the target primary antibody with desired dilution (anti-ACE-2, 1:1000	anti-ACE-2 suggested: None
   For loading, control membranes were probed with anti-β-actin and anti-GAPDH antibodies (1:5000, Cell signaling technology, MA, USA).	anti-β-actin suggested: None anti-GAPDH suggested: None
   The membrane was washed with 1X TBST (3 × 5 min) and then incubated with respective secondary antibodies (Horse anti-mice 1:2000; Horse anti-rabbit 1:2000, (Cell signaling technology, MA, USA) and developed by using ECL chemiluminescent reagent (Bio-Rad, CA, USA)	anti-mice suggested: (AgriSera Cat# AS04 040, RRID:AB_2226396) anti-rabbit suggested: None
   After treatment, the protein-enzyme mixtures were mixed with 2x Laemmli buffer and boiled for 100°C for 5 min and then immunoprobed with anti-Spike protein-specific antibody as described above.	anti-Spike protein-specific suggested: None
   The cells were incubated with anti-S protein Rab (Sino Biological, PA, USA) at 1:1000 in the blocking solution overnight at 4°C, followed by incubation with 1:2000 diluted Alexa Fluor 488 conjugated secondary antibody (Thermo Fisher, MA, USA) for 1 h at room temperature.	anti-S protein Rab suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   , USA), human lung adenocarcinoma cells (A549, a kind gift from Maher Abdulla, Department of Pathology and Microbiology, UNMC and Calu-3, a kind gift from John Dickinson, Internal Medicine Division of Pulmonary, Critical Care and Sleep, UNMC), human normal bronchial epithelial cells (BEAS-2B, a kind gift from Todd Wyatt, Internal Medicine Division of Pulmonary, Critical Care and Sleep, UNMC), human Pancreatic cancer cells (T3M4, a kind gift from Michael A Hollingsworth, Eppley Institute for Research in Cancer, UNMC), and human embryonic kidney epithelial cells (HEK293T, ATCC, VA, USA) were used for this study.	A549 suggested: None Calu-3 suggested: None
   VeroE6, A549, T3M4, and HEK293T were grown in DMEM (Gibco, MA, USA) with 10%	HEK293T suggested: None
   BEAS-2B cells were grown in DMEM with 5% FBS and 1X penicillin and streptomycin.	BEAS-2B suggested: None
   For treatment with VeroE6 cells, bromelain (75	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Software and Algorithms
   Sentences	Resources
   Total hACE2 binding and data analysis were calculated with Bio-Rad ProteOn Manager software (version 3.1).	Bio-Rad ProteOn Manager suggested: None ProteOn suggested: (Proteon Therapeutics, RRID:SCR_004037)
   Statistical Analysis: Statistical analysis was performed with GraphPad Prism version 8 using a two-tailed unpaired t-test.	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 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.
   • 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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2. 

   Version published to 10.1101/2020.09.16.297366 on bioRxiv

   Sep 16, 2020