Fig 1: Set1A participates in the establishment of the mark H3K4me3 on HBV promoters.(A) Human hepatocarcinoma Huh7 cells were transfected with the wild type HBV genome. Set1A recruitment to viral promoters was assayed by ChIP analysis. Immunoprecipitated DNA was quantified by qPCR using specific primers as detailed in Material and Methods. The results are expressed as % of input. The standard deviation was obtained from two PCR reactions and the graph is representative of three independent experiments. (B) Human hepatocarcinoma HepG2 cells were transfected with the wild type HBV genome together with a plasmid encoding for Flag-tagged HBx. Flag-HBx recruitment to viral promoters was assayed by ChIP analysis (left). Then the Flag-HBx immunoprecipitated samples were reChIP with antibodies against Set1A or IgG antibodies as control (right). Immunoprecipitated DNA was quantified by qPCR using specific primers for core promoter. The results are expressed as % of input. (C,D, left) Western blot analyses to monitor overexpression (C) and knock down (D) of Set1A. ?eta actin was used as a loading control. (C,D) H3K4me3 modification was determined by ChIP analysis using specific antibodies from cells overexpressing (C) or knocking down (D) Set1A. Immunoprecipitated DNA was quantified by qPCR using specific primers as detailed in Material and Methods. We normalized the data against the immunoprecipitated H3. The standard deviation was obtained from two PCR reactions and the graphs are representative of three independent experiments.