High content screening identifies cardiac glycoside as an inducer of PML nuclear body formation.
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Abstract
<p><b>A,</b> Scatterplot of the data obtained from a total of 1,008 384-well library plates that were screened, imaged, and analyzed, yielding an average Z’ of 0.50 and a hit rate of 0.3%. IFN 10 U/μl (positive control, dark gray), DMSO (negative control, light gray), compounds (black) <b>B and C,</b> HeLa cells were treated with increasing concentrations of gitoxigenin as well as DMSO as a negative control for 18h. PML NB formation was determined by immunofluorescence with anti-PML antibody and was analyzed using an algorithm described in Materials and Methods. The images are representative PML NB staining of cells treated with DMSO and 0.04 μM, 1.25 μM and 20 μM gitoxigenin for 18h (B). The graphs show fitted curves for PML NB (black diamonds) and cell count (grey circles) over the full dose range of gitoxigenin (C). PML NB values at high concentrations are shown with open diamonds and are excluded from the fitted curve due to the cytotoxicity as evident from dramatic decrease in cell counts at these concentrations.</p