Lipid droplets fuel SARS-CoV-2 replication and production of inflammatory mediators

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1. 

   Version published to 10.1101/2020.08.22.262733v2 on bioRxiv

   Aug 29, 2020
2. 

   ScreenIT

   Aug 24, 2020

   SciScore for 10.1101/2020.08.22.262733: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: Ethics statement: Experimental procedures involving human cells from healthy donors were performed with samples obtained after written informed consent and were approved by the Institutional Review Board (IRB) of the Oswaldo Cruz Foundation/Fiocruz (Rio de Janeiro, RJ, Brazil) under the number 397-07. IRB: Experimental procedures involving human patient cells were performed with samples obtained after written informed consent from all participants or patients’ representatives according to the study protocol approved by the National Review Board (CONEP 30650420.4.1001.0008).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a …

   SciScore for 10.1101/2020.08.22.262733: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: Ethics statement: Experimental procedures involving human cells from healthy donors were performed with samples obtained after written informed consent and were approved by the Institutional Review Board (IRB) of the Oswaldo Cruz Foundation/Fiocruz (Rio de Janeiro, RJ, Brazil) under the number 397-07. IRB: Experimental procedures involving human patient cells were performed with samples obtained after written informed consent from all participants or patients’ representatives according to the study protocol approved by the National Review Board (CONEP 30650420.4.1001.0008).
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	After 2 hours of the plaque, non-adherent cells were washed out and the remaining monocytes were maintained for 24 hours in DMEM containing 5% inactivated male human AB serum (HS; Merck) and 100 U/mL penicillin-streptomycin (P/S; GIBCO) at 37 °C in 5 % CO2.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The cells were incubated with anti-CD14 antibodies (1:10) for 10 min and magnetic beads-conjugates (1:20) for additional 10 min, followed by magnetic recovery of monocytes for 5 min.	anti-CD14 suggested: None
   The purity of human monocytes was above 90 %, as analyzed by flow cytometry analysis (FACScan; Becton Dickinson) using anti-CD3 (BD Biosciences) and anti-CD16 (Southern Biotech) monoclonal antibodies.	anti-CD3 suggested: None anti-CD16 suggested: None
   The double-RNA was labeling by mouse monoclonal antibody J2 clone - Scicons (Schönborn et al., 1991) at 1:500 dilution for overnight, followed by a mouse anti-IgG-Dylight 550 at 1:1000 dilution for 1h.	anti-IgG-Dylight 550 suggested: None
   Membranes were probed overnight with the following antibodies: anti-PPARγ (Santa Cruz Biotechnology, #SC-7196 - H100), anti-CD36 (Proteintech-18836-1-AP), anti-SREBP-1 (Ab-28481), anti-DGAT-1 (Santa Cruz Biotechnology, #SC-271934) and anti-β-actin (Sigma, #A1978).	anti-PPARγ suggested: (Enogene Cat# E2A6073, RRID:AB_2861246) anti-CD36 suggested: None anti-SREBP-1 suggested: None anti-DGAT-1 suggested: None #SC-271934 suggested: (Santa Cruz Biotechnology Cat# sc-271934, RRID:AB_10649947) anti-β-actin suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Human lung epithelial carcinoma cell line (A549 - ATCC/CCL-185) and African green monkey kidney (Vero subtype E6) were cultured in high glucose DMEM supplemented with 10% FBS and 100 U/mL P/S, and were incubated at 37 °C in 5 % CO2.	A549 suggested: None
   The Plaque-forming Assay was performed for virus titration in VERO E6 cells seeded in 24-well plates.	VERO E6 suggested: None
   Lipid droplet staining: Human primary monocytes, A549 cell line, and HMVEC cell line were seeded in coverslips.	HMVEC suggested: None
   Software and Algorithms
   Sentences	Resources
   The numbers of LDs were automatically quantified by ImageJ software analysis from 15 aleatory fields.	ImageJ suggested: (ImageJ, RRID:SCR_003070)

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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
3. 

   Version published to 10.1101/2020.08.22.262733v1 on bioRxiv

   Aug 23, 2020