Differential LIM and LID topology in zebularine-treated cells. LIM and LID sites detected in the range of (tbcg, tQ) in individual cells from Figure 6 (marked in cyan color) are superimposed onto an intermediate optical section in the respective MeC (green) and DAPI (blue) channels (left and middle columns). LIM/LID density curves obtained through morphological erosion of nuclei are shown as cumulative diagrams (right column). These sites correspond with codistribution patterns in Figure 4 for the respective nuclei. The number of graph points in the third column is associated with the number of detected shells; the value of each point refers to the fraction of all sites found in the nucleus up to the next shell. V is the shell volume and Vtot is the total volume of a nucleus. The argument V/Vtot = 0.5 distinguishes all LIM sites that are localized in the peripheral half of the nucleus from LIMs of the interior half (V/Vtot > 0.5). The diagonal line across each plot in represents hypothetically equal density of LIM or LID sites across the nuclear volume. Similar degrees of high rim-like LIM and LID densities are apparent in untreated prostate and hepatic cancer cell nuclei. LIM sites quasi-linearly expand towards the nuclear interior upon the increase of zebularine concentration, with ZEB at 500–1000 μM show large coverage in Huh-7 nuclei and nearly full coverage throughout DU145 nuclei. Also LIDs show an increased distribution in treated cells versus naïve cells, but the changes are more similar across all the applied ZEB and AZA concentrations in both cell types, with most increases occurring in the exterior shells of the nuclei.
Gertych et al. BMC Pharmacology and Toxicology 2013 14:11 doi:10.1186/2050-6511-14-11