Non-permissive SARS-CoV-2 infection in human neurospheres
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SciScore for 10.1101/2020.09.11.293951: (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 Plaque numbers were scored in at least 3 replicates per dilution by independent readers blinded to the experimental group and the virus titers were determined by plaque-forming units (PFU) per milliliter. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The primary antibody was incubated overnight at 4°C [anti-double-stranded RNA (dsRNA) monoclonal antibody (1:200, Scicons)]. anti-double-stranded RNA ( dsRNA )suggested: NoneThen, cells were incubated with primary antibody anti-dsRNA (1:200) overnight at 4°C. anti-dsRNAs…SciScore for 10.1101/2020.09.11.293951: (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 Plaque numbers were scored in at least 3 replicates per dilution by independent readers blinded to the experimental group and the virus titers were determined by plaque-forming units (PFU) per milliliter. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources The primary antibody was incubated overnight at 4°C [anti-double-stranded RNA (dsRNA) monoclonal antibody (1:200, Scicons)]. anti-double-stranded RNA ( dsRNA )suggested: NoneThen, cells were incubated with primary antibody anti-dsRNA (1:200) overnight at 4°C. anti-dsRNAsuggested: (Millipore Cat# MABE1134, RRID:AB_2819101)Day after, cells were incubated with the secondary antibody goat anti-mouse Alexa Fluor 488 (1:400) for 1h. anti-mousesuggested: NoneThen, membranes were incubated with peroxidase-conjugated antibody goat anti-Mouse IgG (H+L), HRP-conjugate (1:10,000, G21040 −Molecular Probes). anti-Mouse IgGsuggested: (Thermo Fisher Scientific Cat# G-21040, RRID:AB_2536527)Experimental Models: Cell Lines Sentences Resources Plaque forming unit assay: For virus titration, monolayers of Vero E6 cells (2 x 104 cell/well) in 96-well plates were infected with serial dilutions of supernatants containing SARS-CoV-2 for 1 hour at 37°C. Vero E6suggested: None, extract samples (40 μg/lane for NP and 15 μg/lane for Vero cells) were separated by electrophoresis on a 10% SDS polyacrylamide gel and transferred to polyvinylidene difluoride (PVDF) membranes. Verosuggested: NoneExperimental Models: Organisms/Strains Sentences Resources SARS-CoV-2 propagation: SARS-CoV-2 obtained from a nasopharyngeal swab from a confirmed case in Rio de Janeiro, Brazil ( SARS-CoV-2suggested: NoneSoftware and Algorithms Sentences Resources Cell cultivation: Human induced pluripotent stem (iPS) cells were obtained from the Coriell Institute for Medical Research repository (GM23279A) or produced in house with CytoTune™-iPS 2.0 Sendai Reprogramming Kit (A16517-Invitrogen) from skin fibroblasts [68] or urine epithelial cells [ CytoTune™-iPSsuggested: NonePrimers, probes, and cycling conditions recommended by the Centers for Disease Control and Prevention (CDC) protocol were used to detect the SARS-CoV-2 (https://www.fda.gov). https://www.fda.govsuggested: (U.S. Food and Drug Administration, RRID:SCR_012945)Cytokine multiplex assay and LDH cytotoxicity assay: A multiplex biometric immunoassay containing fluorescent dyed microbeads was used to measure cytokines in the cell culture supernatant (Bio-Rad Laboratories, Hercules, CA, USA). Bio-Rad Laboratoriessuggested: (Bio-Rad Laboratories, RRID:SCR_008426)Statistics were performed using GraphPad Prism software version 8. GraphPad Prismsuggested: (GraphPad Prism, RRID:SCR_002798)SARS-CoV-2 strains sequence analyses: Nucleotide sequences from our strain (GenBank accession No. MT710714), from strains used in others works [20,37], and from SARS-CoV-2 reference genome (Wuhan-Hu-1, GenBank accession No. NC_045512.2) were retrieved from NCBI database and from Multiple Sequence Alignment (MSA). NCBIsuggested: (NCBI, RRID:SCR_006472)Visualization of SARS-CoV-2 sequences was performed using ClustalW [75] implemented in MEGA X program-version 10.1.8 [76]. ClustalWsuggested: (ClustalW, RRID:SCR_017277)MEGA Xsuggested: NoneResults 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 did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- No funding statement was detected.
- No protocol registration statement was detected.
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