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

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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?

RecJf (D) Signup and view all the answers

What is the purpose of circularizing the digested fragments?

To prevent degradation of the fragments (C) Signup and view all the answers

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

To ligate DNA fragments (C) Signup and view all the answers

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

Times when cell supernatants were not measured (A) Signup and view all the answers

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

HepG2-NTCP cells (C) Signup and view all the answers

What does cccDNA stand for in the given context?

Covalently Closed Circular DNA (B) Signup and view all the answers

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

Data from independent experiments (D) Signup and view all the answers

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

Setup 1 (A) Signup and view all the answers

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

Hepatitis B Core protein (D) Signup and view all the answers

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

It declined at a rate consistent with cccDNA loss with mitosis (A) Signup and view all the answers

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

ELISA (C) Signup and view all the answers

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

Decrease (D) Signup and view all the answers

What does MyrB represent in the context of the study?

Treatment with a virus inhibitor (C) Signup and view all the answers

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

To observe the effect of cccDNA loss with mitosis (C) Signup and view all the answers

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

Dilution model (A) Signup and view all the answers

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

Whether cells were lysed immediately after cell mitosis or cultured until 12 dpi. (A) Signup and view all the answers

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

Different levels of cccDNA decrease per passage. (D) Signup and view all the answers

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

Partial loss of cccDNA during mitosis. (A) Signup and view all the answers

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

Amplification of cccDNA following mitosis. (D) Signup and view all the answers

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

Other cccDNA degradation mechanisms like cytokine-induced degradation. (A) Signup and view all the answers

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

HepG2-NTCP and HepaRG-NTCP cells. (C) Signup and view all the answers

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

They were viable and expressed no new viral antigens. (B) Signup and view all the answers

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

Produce two uninfected daughter cells. (B) Signup and view all the answers

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

tRFP mRNA (B) Signup and view all the answers

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

~100,000 cells per cm2 (B) Signup and view all the answers

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

tRFP expression was stable. (C) Signup and view all the answers

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

New tRFP was not expressed in the daughter cells. (A) Signup and view all the answers

Ilastik software

Software used for image editing in the study.

NucleoSpin Tissue kit

Kit used for extracting total cellular DNA.