FIG 10.

Cell-type-specific effects of ganciclovir on TB40-ANCHOR3 HCMV infection and IC₅₀ determination by automated high-content imaging. (a) TB40-ANCHOR3 HCMV-infected cells were treated or not with 12.5 μM ganciclovir; 72 h p.i., the cells were observed using a Thermo Scientific Cellomics Arrayscan Vti microscope, and the images were analyzed with the compartmental analysis algorithm, allowing detection of viral DNA (red) (see Materials and Methods for an explanation). (b) Experiment similar to that shown in panel a, but TB40-ANCHOR3 HCMV-infected cells were treated with increasing doses of ganciclovir. The results of the quantification were plotted against ganciclovir concentrations (see the text), allowing precise determination of the IC₅₀ and IC₉₀. Two identical experiments were performed on separate plates (A and B). (c) Time course of TB40-ANCHOR3 HCMV infections in MRC5 or ARPE-19 cells in the presence or absence of 2.5 μM ganciclovir. In MRC5 cells, infection progressed to more than 70% infected cells 10 days p.i., and the infection was partly controlled by ganciclovir. ARPE-19 cells did not seem to be highly permissive, and only 2 to 3% of the cells became infected in the presence or absence of ganciclovir. (d) Same as panel c but using viral DNA quantification as a readout. d, days. The error bars represent SD.