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HepG2-NTCP Cells Infected with WT HBV vs Replication-Deficient Mutants
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HepG2-NTCP Cells Infected with WT HBV vs Replication-Deficient Mutants

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Questions and Answers

What software was used for image editing?

  • Ilastik (correct)
  • Photoshop
  • GIMP
  • ImageMagick

    • Which kit was used for extracting total cellular DNA?

  • NucleoSpin® Tissue kit (correct)
  • Quick-DNA Miniprep kit
  • GeneJET Genomic DNA Purification Kit
  • QIAamp DNA Mini kit

    • What is the purpose of the restriction digestion reaction?

  • To extract DNA
  • To label DNA
  • To amplify DNA
  • To digest DNA into fragments (correct)

    • Which enzyme was used in the restriction digestion reaction?

    <p>RecJf</p> Signup and view all the answers

    What is the purpose of circularizing the digested fragments?

    <p>To prevent degradation of the fragments</p> Signup and view all the answers

    What is the purpose of adding T4 DNA Ligase in the reaction?

    <p>To ligate DNA fragments</p> Signup and view all the answers

    What is represented by the dashed black lines in Fig. 3B?

    <p>Times when cell supernatants were not measured</p> Signup and view all the answers

    Which cell type was infected with a Gaussia luciferase-expressing reporter HBV?

    <p>HepG2-NTCP cells</p> Signup and view all the answers

    What does cccDNA stand for in the given context?

    <p>Covalently Closed Circular DNA</p> Signup and view all the answers

    What do the light lines connecting data points in Fig. 2A represent?

    <p>Data from independent experiments</p> Signup and view all the answers

    In which setup were HepG2-NTCP cells serially split after infection?

    <p>Setup 1</p> Signup and view all the answers

    What does HBc refer to in the context of the experiment?

    <p>Hepatitis B Core protein</p> Signup and view all the answers

    What was found regarding the protein expression from cccDNA in relation to cccDNA copy number?

    <p>It declined at a rate consistent with cccDNA loss with mitosis</p> Signup and view all the answers

    Which technique was found to be insufficiently sensitive to detect low levels of secretion from cccDNA?

    <p>ELISA</p> Signup and view all the answers

    What was the effect observed on the number of HBc-positive cells with each additional split?

    <p>Decrease</p> Signup and view all the answers

    What does MyrB represent in the context of the study?

    <p>Treatment with a virus inhibitor</p> Signup and view all the answers

    What was the purpose of visualizing HBV-infected HepG2-NTCP cells using fluorescence microscopy?

    <p>To observe the effect of cccDNA loss with mitosis</p> Signup and view all the answers

    Which model is used to describe the expected number of HBc-positive cells following mitosis?

    <p>Dilution model</p> Signup and view all the answers

    What was the main difference between setup 1 (green) and setup 2 (purple) in the experimental setups?

    <p>Whether cells were lysed immediately after cell mitosis or cultured until 12 dpi.</p> Signup and view all the answers

    What do the top and bottom dashed lines in the dilution and loss models predict?

    <p>Different levels of cccDNA decrease per passage.</p> Signup and view all the answers

    What does the area between the dilution and loss models represent?

    <p>Partial loss of cccDNA during mitosis.</p> Signup and view all the answers

    What do higher levels of cccDNA compared to the dilution model suggest?

    <p>Amplification of cccDNA following mitosis.</p> Signup and view all the answers

    What is represented by levels lower than the loss model in terms of cccDNA post-mitosis?

    <p>Other cccDNA degradation mechanisms like cytokine-induced degradation.</p> Signup and view all the answers

    In what type of cells were the experiments conducted involving setup 1 and setup 2?

    <p>HepG2-NTCP and HepaRG-NTCP cells.</p> Signup and view all the answers

    What was observed about daughter cells of HBV-infected hepatocytes after mitosis?

    <p>They were viable and expressed no new viral antigens.</p> Signup and view all the answers

    What was the fate of individual HBV-infected cells that undergo mitosis?

    <p>Produce two uninfected daughter cells.</p> Signup and view all the answers

    What was used to track living HBV-infected cells in the study?

    <p>tRFP mRNA</p> Signup and view all the answers

    What was the concentration of HepG2-NTCP cells infected with tRFP-HBV seeded in chamber slides?

    <p>~100,000 cells per cm2</p> Signup and view all the answers

    What was observed in high-confluence areas when it came to tRFP expression in non-dividing cells?

    <p>tRFP expression was stable.</p> Signup and view all the answers

    What was the outcome after photo-bleaching all existing tRFP and allowing the cells to recover?

    <p>New tRFP was not expressed in the daughter cells.</p> Signup and view all the answers

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