Figure 4
Comparative analyses of histone acetylation and chromatin structure in met murine hepatocytes (MMH)-D3 versus hepatitis B virus met murine hepatocytes (HBV-Met) cells. A. Western blot analyses of H3K9ac (left) or H2A.Zac (right) enrichment in MMH-D3 and HBV-Met cells. Beta-actin was used as loading control. B-D. Assays of histone acetyltransferase (HAT)/ histone deacetylase (HDAC) activities. B. HAT activity ELISA using nuclear extracts from 10.000 of each of both cell lines MMH-D3 and HBV-Met. The dots and the trend line correspond to the kit’s standards, whereas the boxplots (referring to the y-axis only) represent the results of several measurements (n =10 for each cell line) using MMH-D3 or HBV-Met cells. C. Results of class I/II HDAC activity measurements over time using 10.000 cells from each of the two cell lines for luminescence reporter assays. D. Results of class III HDAC activity (sirtuins) measurements over time using 20.000 cells from each of the two cell lines for luminescence reporter assays. E. Results of MNase digests over time using isolated nuclei from 1x 106 cells per time point. F. Fold-change differences of H2A.Zac/H3K9ac recovery at selected sites. HBV-Met versus MMH-D3 or HBV-Met with suppressed hepatitis B virus (HBV) replication. For each site recovery from HBV-Met was used for normalization.