The Inhaled Steroid Ciclesonide Blocks SARS-CoV-2 RNA Replication by Targeting the Viral Replication-Transcription Complex in Cultured Cells

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1. 

   ScreenIT

   Mar 1, 2021

   SciScore for 10.1101/2020.08.22.258459: (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 cells were then incubated with a mixture of rabbit anti-SARS-nsp4 (1:500; ab181620; Abcam, USA) and mouse anti-dsRNA (1:1000; J2-1709; Scicons, Hungary) antibodies for 1 h at RT, washed three times with PBS, and incubated for 1 h at RT with a mixture of Alexa Fluor 594 conjugated anti-rabbit IgG (1:500; A11012; ThermoFisher, USA) and Alexa Fluor 488-conjugated anti-mouse IgG (1:500; A10680; ThermoFisher, USA).	anti-SARS-nsp4 suggested: None anti-dsRNA sugg…

   SciScore for 10.1101/2020.08.22.258459: (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 cells were then incubated with a mixture of rabbit anti-SARS-nsp4 (1:500; ab181620; Abcam, USA) and mouse anti-dsRNA (1:1000; J2-1709; Scicons, Hungary) antibodies for 1 h at RT, washed three times with PBS, and incubated for 1 h at RT with a mixture of Alexa Fluor 594 conjugated anti-rabbit IgG (1:500; A11012; ThermoFisher, USA) and Alexa Fluor 488-conjugated anti-mouse IgG (1:500; A10680; ThermoFisher, USA).	anti-SARS-nsp4 suggested: None anti-dsRNA suggested: (Millipore Cat# MABE1134, RRID:AB_2819101) anti-rabbit IgG suggested: (Molecular Probes Cat# A-11012, RRID:AB_141359) anti-mouse IgG suggested: (Thermo Fisher Scientific Cat# A-10680, RRID:AB_2534062)
   Experimental Models: Cell Lines
   Sentences	Resources
   Cells and viruses: Hep-2, HeLa229, MDCK, Calu-3, Vero, Vero/TMPRSS2 and VeroE6/TMPRSS2 cells were maintained in Dulbecco’s modified Eagle medium high glucose (DMEM, Sigma-Aldrich, USA), and DBT cells were maintained in DMEM (Nissui, Japan), supplemented with 5% fetal bovine serum (Gibco-BRL, USA).	Hep-2 suggested: None MDCK suggested: CLS Cat# 602280/p823_MDCK_(NBL-2, RRID:CVCL_0422) Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059) VeroE6/TMPRSS2 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   BHK-21 cells were grown in a single well of a six-well plate in 10% FCS-MEM and transfected with 3 μg of BAC plasmid with Lipofectamine 3,000 (Thermo Fisher, USA)	BHK-21 suggested: None
   The supernatants were collected and propagated once using Vero/TMPRSS2 cells.	Vero/TMPRSS2 suggested: JCRB Cat# JCRB1818, RRID:CVCL_YQ48)
   Vero and VeroE6/TMPSS2 cells were used to passage MERS-CoV and SARS-CoV-2, respectively.	VeroE6/TMPSS2 suggested: None
   Software and Algorithms
   Sentences	Resources
   Indexed libraries were then converted and sequenced (150-bp paired-end reads) using the DNBSEQ-G400 (MGI Tech., Shenzhen, China; operated by GENEWIZ, South Plainfield, NJ, USA).	GENEWIZ suggested: (GENEWIZ, RRID:SCR_003177)

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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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   Read the original source
2. Version published to 10.1128/jvi.01648-20

   Dec 9, 2020
3. 

   Version published to 10.1101/2020.08.22.258459 on bioRxiv

   Aug 24, 2020