The SARS-CoV-2 spike protein alters barrier function in 2D static and 3D microfluidic in-vitro models of the human blood–brain barrier

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1. Version published to 10.1016/j.nbd.2020.105131

   Dec 1, 2020
2. 

   ScreenIT

   Jun 17, 2020

   SciScore for 10.1101/2020.06.15.150912: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: Healthy tissue was provided (under informed consent) by the Laboratory of Developmental Biology (University of Washington, Seattle, WA) with approval granted by Temple University’s (Philadelphia, PA) Institutional Review Board and in full compliance by the National Institutes of Health’s (NIH) ethical guidelines. IRB: Healthy tissue was provided (under informed consent) by the Laboratory of Developmental Biology (University of Washington, Seattle, WA) with approval granted by Temple University’s (Philadelphia, PA) Institutional Review Board and in full compliance by the National Institutes of Health’s (NIH) ethical guidelines.
   Randomization	not detected.
   Bli…

   SciScore for 10.1101/2020.06.15.150912: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: Healthy tissue was provided (under informed consent) by the Laboratory of Developmental Biology (University of Washington, Seattle, WA) with approval granted by Temple University’s (Philadelphia, PA) Institutional Review Board and in full compliance by the National Institutes of Health’s (NIH) ethical guidelines. IRB: Healthy tissue was provided (under informed consent) by the Laboratory of Developmental Biology (University of Washington, Seattle, WA) with approval granted by Temple University’s (Philadelphia, PA) Institutional Review Board and in full compliance by the National Institutes of Health’s (NIH) ethical guidelines.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The sections were subsequently incubated in rabbit anti-human ACE2 antibody (1:500, Abcam, ab15348) for 1 hr at RT.	anti-human ACE2 suggested: (Abcam Cat# ab15348, RRID:AB_301861)
   Positive antibody binding was detected using anti-rabbit IgG ImmPRESS-AP polymer reagent (Vector Laboratories, MP-5401) and visualized via a 10 min incubation in Vector Blue AP substrate (Vector Laboratories, SK-5300).	anti-rabbit IgG suggested: (Vector Laboratories Cat# MP-5401, RRID:AB_2336536) MP-5401 suggested: (Vector Laboratories Cat# MP-5401, RRID:AB_2336536)
   Cells were resuspended for 30 min in 100 μL of flow cytometry buffer and anti-ICAM-1 (Pe-Cy7, Biolegend) and anti-VCAM (APC, Biolegend) preconjugated primary antibodies.	anti-ICAM-1 suggested: None anti-VCAM (APC suggested: None
   Blocked protein blot was incubated with affinity-purified rabbit anti-ACE2 (1:1000, Abcam, Cat No 15348), and mouse anti-b-actin (1:5000, Sigma, Cat No A5441), in PBS with 0.05% Tween-20 and 10% Odyssey blocking buffer at 4°C overnight, followed by incubation with goat anti-rabbit IRDye 800CW IgG and goat anti-mouse IRDye 680RD secondary antibody in PBS (1:20,000) at room temperature for 1 h.	anti-ACE2 suggested: None anti-b-actin suggested: (Sigma-Aldrich Cat# A5441, RRID:AB_476744) anti-rabbit suggested: (LI-COR Biosciences Cat# 925-68073, RRID:AB_2716687) anti-mouse suggested: None
   Software and Algorithms
   Sentences	Resources
   Immunohistochemistry and Imaging: Formalin-fixed paraffin-embedded human frontal cortex brain tissue blocks of both normal and diseased origin (mixed-type dementia, cardiovascular and pulmonary dysfunction) were procured from ProteoGenex, Inc. (Inglewood, CA), and serially sectioned at a thickness of 5 µm each.	ProteoGenex suggested: (ProteoGenex, RRID:SCR_013844)
   For Imaging, all sections were scanned with the Aperio AT2 slide scanner (Leica Biosystems) and analyzed using Aperio ImageScope software (v12.3.2.8013).	ImageScope suggested: (ImageScope, RRID:SCR_014311)
   10,000 events per sample were acquired with a FACS BD Canto II flow cytometer (BD Biosciences) and data was then analyzed with FlowJo software (Tree Star, Ashland, OR, USA).	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Band intensities were quantified using ImageJ software (NIH, Bethesda).	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   The data collected was analyzed using Prism v6.0 (GraphPad Software, San Diego, CA).	Prism suggested: (PRISM, RRID:SCR_005375) 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 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 page 34. 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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3. 

   Version published to 10.1101/2020.06.15.150912v1 on bioRxiv

   Jun 15, 2020