Decitabine Reactivation of FoxM1-Dependent Endothelial Regeneration and Vascular Repair for Potential Treatment of Elderly ARDS and COVID-19 Patients

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   ScreenIT

   May 2, 2021

   SciScore for 10.1101/2021.04.29.442061: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: The animal care and study protocols were approved by the Institutional Animal Care and Use Committees of Northwestern University and The University of Illinois at Chicago. IRB: Human specimens: All experiments with human tissue samples were performed under protocols approved by the Institutional Review Boards at the University of Chicago and Ann & Robert H.
   Sex as a biological variable	Both male and female mice were used in the experiments.
   Randomization	Fifteen 63X fields each section were randomly selected and examined.
   Blinding	not detected.
   Power Analysis	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Mouse lung cryosections were stained overnight with anti-BrdU (1:3, BD …

   SciScore for 10.1101/2021.04.29.442061: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: The animal care and study protocols were approved by the Institutional Animal Care and Use Committees of Northwestern University and The University of Illinois at Chicago. IRB: Human specimens: All experiments with human tissue samples were performed under protocols approved by the Institutional Review Boards at the University of Chicago and Ann & Robert H.
   Sex as a biological variable	Both male and female mice were used in the experiments.
   Randomization	Fifteen 63X fields each section were randomly selected and examined.
   Blinding	not detected.
   Power Analysis	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Mouse lung cryosections were stained overnight with anti-BrdU (1:3, BD Biosciences, or 1:1000 Cell Signaling Technology, Danvers, MA) and incubated with Alexa Fluo 488-conjugated secondary antibody (1:200, Thermal Fisher Scientific, Waltham, MA)	anti-BrdU suggested: None
   Lung vascular ECs were immunostained with anti-vWF (1:300, Sigma-Aldrich, St Louis, MO) and anti-CD31 (1:100, BD Bioscience) antibodies at 4°C.	anti-vWF suggested: None
   Western blot analysis was performed using an anti-FoxM1 antibody (1:800, sc-376471, Santa Cruz Biotechnology, Santa Cruz, CA) and the same blot was incubated with anti-β-actin antibody (1:3000, BD Biosciences) as a loading control.	anti-FoxM1 suggested: (Santa Cruz Biotechnology Cat# sc-376471, RRID:AB_11150135) sc-376471 , Santa Cruz Biotechnology , Santa Cruz , CA suggested: None anti-β-actin suggested: None
   After RNAscope assay, the slides were incubated in blocking buffer (3% BSA, 1% FBS and 0.1% normal donkey serum) for 1 h followed by with a primary antibody against CD31 (Cat # Ab28364, Abcam, Cambridge, MA) at 4°C overnight.	CD31 suggested: (Abcam Cat# ab28364, RRID:AB_726362)
   The sections were washed and incubated with appropriate anti-rabbit secondary antibody labeled with Alexa Fluor 488 for 1 h.	anti-rabbit suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Foxm1 transgenic (Foxm1Tg) mice driven by the Rosa26 promoter were described previously (35).	Foxm1Tg suggested: None
   C57BL/6 WT mice (2 mos.	C57BL/6 suggested: None
   At 3h following irradiation, the mice were transplanted with 10 million of bone marrow cells (200 μl of EBM2 medium) freshly isolated from EndoSCL-CreERT2 transgenic mice (2 mos. old) through tail vein injection.	EndoSCL-CreERT2 suggested: None
   Software and Algorithms
   Sentences	Resources
   The nuclei were counterstained with DAPI (Thermal Fisher Scientific).	Thermal Fisher Scientific suggested: None
   Statistical significance was determined by one-way ANOVA with a Dunnett post hoc analysis that calculates P values corrected for multiple comparisons for equal variance or Kruskal-Wallis test for unequal variance using Prism 7 (Graphpad Software, Inc. La Jolla, CA)	Prism suggested: (PRISM, RRID:SCR_005375) Graphpad suggested: (GraphPad, RRID:SCR_000306)

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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
   NCT04482621	Recruiting	Decitabine for Coronavirus (COVID-19) Pneumonia- Acute Respi…

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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: Please consider improving the rainbow (“jet”) colormap(s) used on pages 28 and 44. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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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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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2021.04.29.442061v1 on bioRxiv

   Apr 30, 2021