Bromodomain and Extraterminal Protein Inhibitor, Apabetalone (RVX-208), Reduces ACE2 Expression and Attenuates SARS-Cov-2 Infection In Vitro

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1. Version published to 10.3390/biomedicines9040437

   Apr 18, 2021
2. 

   ScreenIT

   Mar 15, 2021

   SciScore for 10.1101/2021.03.10.432949: (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
   Membranes were blocked and incubated with anti-ACE2 (R&D Systems AF933) or anti-GAPDH (Cell Signaling 5174) antibodies overnight at 4°C, followed by incubation with corresponding HRP-conjugated secondary antibodies (Bio-Rad or Abcam) for 1 h at room temperature.	anti-ACE2 suggested: None anti-GAPDH (Cell Signaling 5174) suggested: (Cell Signaling Technology Cat# 5174, RRID:AB_10622025)
   Actin was stained with an anti-β-ACTIN antibody conjugated directly to …

   SciScore for 10.1101/2021.03.10.432949: (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
   Membranes were blocked and incubated with anti-ACE2 (R&D Systems AF933) or anti-GAPDH (Cell Signaling 5174) antibodies overnight at 4°C, followed by incubation with corresponding HRP-conjugated secondary antibodies (Bio-Rad or Abcam) for 1 h at room temperature.	anti-ACE2 suggested: None anti-GAPDH (Cell Signaling 5174) suggested: (Cell Signaling Technology Cat# 5174, RRID:AB_10622025)
   Actin was stained with an anti-β-ACTIN antibody conjugated directly to peroxidase (Sigma A3854).	anti-β-ACTIN suggested: None
   The cells were then washed and incubated with PE-conjugated goat anti-human Fc antibodies (Thermo Fisher 12-4998-82) for 30 min at 4°C.	anti-human Fc suggested: (Thermo Fisher Scientific Cat# 12-4998-82, RRID:AB_465926)
   The cells were incubated with anti-Spike protein Rab (Sino Biological) at 1:1000 in blocking solution overnight at 4°C, followed by incubation with 1:2000 diluted Alexa Fluor 488 conjugated secondary antibody (Thermo Fisher) for 1 h at room temperature.	anti-Spike protein Rab (Sino Biological) suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Human bronchial epithelial Calu-3 cells and African green monkey kidney epithelial Vero E6 cells (ATCC) were maintained in complete medium (Eagle’s Minimum Essential Medium [EMEM, ATCC]) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin and eptomycin (P/S).	Calu-3 suggested: None
   HepG2 (ATCC) and Huh-7 (JCRB Cell Bank) were cultured in medium recommended by the suppliers.	HepG2 suggested: CLS Cat# 300198/p2277_Hep-G2, RRID:CVCL_0027) Huh-7 suggested: CLS Cat# 300156/p7178_HuH7, RRID:CVCL_0336)
   Human Karpas-299 T cells (a kind gift from Dr. Iqbal, UNMC; originally from DMSZ) were cultured in RPMI-1640 supplemented with 10% FBS and P/S.	Karpas-299 suggested: ECACC Cat# 06072604, RRID:CVCL_1324)
   Immunoblot Analysis: Calu-3 or Vero E6 cells treated with BETi for 48 hours were lysed and sonicated as previously described 31.	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   ImageJ or Quantity 4.6.9 (Bio-Rad) software were used for densitometric quantification of bands in immunoblots 32.	ImageJ suggested: (ImageJ, RRID:SCR_003070) Quantity suggested: None
   Cell surface ACE2 protein levels were measured on a BD FACSCelesta (BD Biosciences) and analyzed with FlowJo version 10 software (BD Biosciences).	FlowJo suggested: (FlowJo, RRID:SCR_008520)

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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: We found the following clinical trial numbers in your paper:
   

   Identifier	Status	Title
   NCT04353284	Recruiting	Camostat Mesylate in COVID-19 Outpatients
   NCT04524663	Recruiting	Oral Camostat Compared With Standard Supportive Care in Mild…
   NCT04470544	Recruiting	Camostat Mesilate Treating Patients With Hospitalized Patien…
   NCT04542213	Recruiting	Dipeptidyl Peptidase-4 Inhibitor (DPP4i) for the Control of …

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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
3. 

   Version published to 10.1101/2021.03.10.432949v1 on bioRxiv

   Mar 11, 2021