*: The reduction in levels of HBV genome-containing nucleocapsids was compared with that of core proteins and found to be highly significant (P < 0.05 as monitored by Pearson χ 2 test). Results shown are representative of three independent experiments. The signals including 3.5-kb RNA, core proteins and genome-containing nucleocapsids were quantified by densitometry analysis and expressed as the average percentage of respective control cells from three independent experiments to indicate the inhibitory effect of IL-6. 17 In general, it allows one to determine whether specific nucleotide sequences in a cloned probe are present in a sample of genomic DNA. HBV genome-containing nucleocapsids were then detected by Southern blot analysis of the disrupted nucleocapsids using an HBV-specific probe (panel for Genome-containing nucleocapsids). One of the most useful techniques for analyzing a gene at the level of genomic DNA is Southern blotting, named for its originator, E.M. For particle blot analysis, equal amounts of cell lysates were assayed for viral capsid by native agarose gel electrophoresis. Southern blot filters were first hybridized to the ETS2 probe, freed of the probe, and hybridized to the reference 0.45-kb probe. dard Southern blotting, we had to use densitometry analysis to. The expression level of β-actin was used as an internal control for sample loading. successively hybridized with the probes tested and with the non-X chromosome rDNA probe. The total core protein was subsequently detected by immunoblot analysis with antibodies against core protein. For Western blot analysis (panel for Proteins), cell lysates were harvested and equal amounts of samples were separated by SDS-polyacrylamide gel electrophoresis. The expression level of 18S rRNA was used as an internal control for sample loading. For RNA analysis (panel for RNA), total RNAs were extracted and equal amounts of samples were subjected to Northern blot analysis to reveal the expression profile of the HBV transcripts, namely those of 3.5-kb and 2.4-kb/2.1-kb respectively. Stain-free western blotting allows you to quickly check electrophoresis and blot transfer quality and obtain truly quantitative western blotting results. (A) SB using HBV DNA probes (top panel) and densitometric analysis of the. Confluent cells (day0) were treated with or without 20 ng/ml of IL-6 for 2 days (day2), 4 days (day4) or 6 days (day6). Southern blot (SB) permits cccDNA visualisation but lacks sensitivity and is. IL-6 decreases the levels of HBV transcripts, core protein, and genome-containing nucleocapsids.
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