WIN 55,212-2 shows anti-inflammatory and survival properties in human iPSC-derived cardiomyocytes infected with SARS-CoV-2

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1. 

   Version published to 10.1101/2021.02.20.431855v3 on bioRxiv

   Sep 17, 2021
2. 

   Version published to 10.1101/2021.02.20.431855v2 on bioRxiv

   Jul 19, 2021
3. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2021.02.20.431855: (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	Plaque numbers were scored in at least 3 replicates per dilution by independent readers blinded to the experimental group and the virus titers were determined by plaque-forming units (PFU) per milliliter. 2.5.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cardiomyocytes were incubated in primary antibodies diluted in a blocking buffer at 4° overnight (anti-SARS-CoV-2 convalescent serum from a positive COVID-19 patient (1:1000) and anti-cardiac troponin T (TNNT2) (1:500, MA5-12960 - Invitrogen).	anti-SARS-CoV-2 sugges…

   SciScore for 10.1101/2021.02.20.431855: (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	Plaque numbers were scored in at least 3 replicates per dilution by independent readers blinded to the experimental group and the virus titers were determined by plaque-forming units (PFU) per milliliter. 2.5.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Cardiomyocytes were incubated in primary antibodies diluted in a blocking buffer at 4° overnight (anti-SARS-CoV-2 convalescent serum from a positive COVID-19 patient (1:1000) and anti-cardiac troponin T (TNNT2) (1:500, MA5-12960 - Invitrogen).	anti-SARS-CoV-2 suggested: None anti-cardiac troponin T (TNNT2 suggested: (Hytest Cat# RC4T19-RecChim406, RRID:AB_2889127)
   Next, cardiomyocytes were incubated with the secondary antibody diluted in a blocking buffer: goat anti-Human Alexa Fluor 647 (1:400; A-21445 - Invitrogen) and goat anti-Mouse 594 (1:400; A-11032 - Invitrogen) for 1h.	anti-Human Alexa Fluor 647 suggested: (Molecular Probes Cat# A-21445, RRID:AB_2535862)
   Membranes were incubated overnight at 4°C with primary antibodies anti-ACE2 (1: 1000; MA5-32307 - Thermo Fisher), anti-ACTIN (1: 2000; MAB1501, Millipore) diluted in TBS-T with 5% non-fat milk and anti-CB1 (1:500; SC-10066, Santa Cruz).	anti-ACE2 suggested: (Thermo Fisher Scientific Cat# MA5-32307, RRID:AB_2809589) anti-ACTIN suggested: (Millipore Cat# MAB1501, RRID:AB_2223041) anti-CB1 suggested: (Santa Cruz Biotechnology Cat# sc-10066, RRID:AB_637711) SC-10066 suggested: None
   Membranes were washed to be incubated with peroxidase-conjugated antibodies IgG (H + L), HRP-conjugate: goat anti-mouse (1: 10.000, G21040, Molecular Probes) goat anti-rabbit (1: 10.000, G21234, Molecular Probes) and rabbit anti-goat (1: 2.000, 61-1620, Invitrogen).	anti-mouse suggested: None anti-rabbit suggested: (Innovative Research Cat# G-21234, RRID:AB_1500696) anti-goat suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   SARS-CoV-2 titration: For virus titration, monolayers of Vero E6 cells (2 × 104 cell/well) in 96-well plates were infected with serial dilutions of supernatants containing SARS-CoV-2 for 1 hour at 37°C.	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Cytokine multiplex assay and LDH cytotoxicity assay: A multiplex biometric immunoassay containing fluorescent dyed microbeads was used for plasma cytokine measurement (Bio-Rad Laboratories, Hercules, CA, USA).	Bio-Rad Laboratories suggested: (Bio-Rad Laboratories, RRID:SCR_008426)
   Next, membranes were blocked again and proceeded with the above-described steps. 2.11. Statistics: Statistical analyses were performed using GraphPadPrism software version 8.0 (GraphPad, EUA).	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 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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   Read the original source
4. 

   Version published to 10.1101/2021.02.20.431855v1 on bioRxiv

   Feb 21, 2021