Degradation of SARS-CoV-2 receptor ACE2 by the E3 ubiquitin ligase Skp2 in lung epithelial cells
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SciScore for 10.1101/2020.10.13.337774: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Patient samples: The study was approved by the research ethics committees of the Chinese Academy of Medical Sciences Cancer Institute and Hospital.
IACUC: All animal studies were conducted according to protocols approved by the Animal Ethics Committee of our hospital.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 lung biopsy samples of patients with benign diseases (Supplementary Table 1) were obtained from Department of Pathology, Peking Union Medical College Hospital, and were analyzed by … SciScore for 10.1101/2020.10.13.337774: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IRB: Patient samples: The study was approved by the research ethics committees of the Chinese Academy of Medical Sciences Cancer Institute and Hospital.
IACUC: All animal studies were conducted according to protocols approved by the Animal Ethics Committee of our hospital.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 lung biopsy samples of patients with benign diseases (Supplementary Table 1) were obtained from Department of Pathology, Peking Union Medical College Hospital, and were analyzed by immunohistochemistry assay using an anti-ACE2 and anti-Skp2 antibodies. anti-ACE2suggested: Noneanti-Skp2suggested: NoneAntibodies and reagents: Antibodies used in this study included rabbit polyclonal anti-human ACE2 (#ab15348, Abcam, Cambridge, MA, USA; 1:200 for immunofluorescence), rabbit monoclonal anti-human ACE2 (#ab108252, Abcam; 1:1000 for Western blot), anti-human ACE2suggested: Nonerabbit polyclonal anti-human ACE2 (#4355, Cell Signaling Technology, Danvers, MA, USA; 1:1000 for Western blot), anti-TMPRSS2 (#ab92323, Abcam; 1:1000 for Western blot), rabbit anti-Furin (#ab183595, Abcam,; 1:1000 for Western blot), rabbit anti-Skp2 (#2652, Cell Signaling Technology; 1:1000 for Western blot), anti-GAPDH (#5174, Cell Signaling Technology; 1:1000 for Western blot), and anti-β-Actin (#A5441, Sigma, St. Louis, MO, USA; 1:5000 for Western blot). anti-Furinsuggested: Noneanti-β-Actinsuggested: (Sigma-Aldrich Cat# A5441, RRID:AB_476744)Experimental Models: Cell Lines Sentences Resources In vitro ubiqutination assay: Ubiquitination assay was performed with Ubiquitylation Assay Kit (abcam; ab139467) using recombinant carrier free ACE2 (# 933-ZN-010) and Skp1/Skp2 (#E3-521-025) proteins bought from R&D Systems (Minneapolis, MN, USA) or Flag-ACE2 and Flag-Skp1/Skp2 purified from 293T cells. 293Tsuggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)To infect 16HBE and 293T-ACE2 cells with pseudovirions, the cells were seeded onto 96-well plates, pre-treated with CSE (10%), BaP (5 μM), or NNK (5 μM) for 48 h, followed by co-incubation with 100 μl media containing pseudovirions for 48 h. 293T-ACE2suggested: RRID:CVCL_YZ65)Experimental Models: Organisms/Strains Sentences Resources A/J mice (5-6 weeks old) were purchased from the Jackson Laboratory (Bar Harbor, Maine, USA), and were bred and maintained in a specific pathogen-free environment. A/Jsuggested: NoneSoftware and Algorithms Sentences Resources Statistical analysis: All statistical analyses were conducted using the software SPSS 26.0 for Windows (Chicago, IL, USA). SPSSsuggested: (SPSS, RRID:SCR_002865)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).
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.Results from TrialIdentifier: No clinical trial numbers were referenced.
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).
Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 27. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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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