%0 Figure %A Feng, Hui %A Chen, Ping %A Zhao, Fei %A Nassal, Michael %A Hu, Kanghong %D 2013 %T In vitro priming activities of selected ε RNAs showing reduced DNA accumulation in cells. %U https://plos.figshare.com/articles/figure/_In_vitro_priming_activities_of_selected_949_RNAs_showing_reduced_DNA_accumulation_in_cells_/777059 %R 10.1371/journal.pone.0072798.g006 %2 https://ndownloader.figshare.com/files/1175806 %K Biochemistry %K Nucleic acids %K rna %K microbiology %K Virology %K Viral replication %K Viral nucleic acid %K Viral packaging %K Viral replication complex %K Viral enzymes %K Gastroenterology and hepatology %K Liver diseases %K Infectious hepatitis %K Hepatitis B %K vitro %K priming %K activities %K rnas %K reduced %K dna %K accumulation %X

(A) Increased priming signals by co-expression of wild-type ε RNA and increased α-32P-dATP concentration. FLAG-tagged HBV P protein from cells transfected with only the P protein vector (lane P), or cotransfected with a wild-type ε RNA expression vector (lanes P + ε) was immobilized on anti-FLAG antibody beads. One third each of the immunoprecipitate was incubated with 1 µl or 2 µl of α-32P-dATP (3,000 Ci/mmol) as reported [44]; subsequently, the beads were boiled in SDS-PAGE sample buffer, and the released material was analyzed by SDS-PAGE and autoradiography. The remaining one third of the immunopellet was analyzed for FLAG-tagged P protein by Western blotting (panel anti-FLAG). (B) In vitro priming activities of selected ε RNA variants. FLAG-tagged P protein complexes with the indicated ε RNAs were expressed, affinity purified and subjected to in vitro priming conditions as in (A), using 2 µl α-32P-dATP. The molecular mass marker positions indicated on the left (in kDa) are approximations inferred from the respective marker protein positions on the SDS-PAGE gels used for the anti-FLAG immunoblots which were run in parallel under identical conditions. Numbers below the autoradiogram indicate mean signal intensities ± standard deviation from two independent experiments relative to that produced by the wild-type ε RNA complex which was set to 100.

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