FIG. 5.

Effect of BFA and cerulenin on GFLV multiplication in infected T-BY2 protoplasts. Immediately after electroporation of the protoplasts with viral RNA, 10 and 30 μg of BFA/ml or 30 and 45 μM cerulenin (final concentrations) were added, and the protoplasts were further incubated for 48 h. Cell viability was assessed on aliquots of protoplasts by adding 0.1% FDA. The remaining protoplasts were fixed and immunolabeled with anti-VPg or anti-CP antibodies and A568 conjugate. The number of infected cells, as revealed by the anti-VPg or anti-CP signal, was counted versus the total number of protoplasts determined under UV illumination after staining the nucleus with Hoechst 33258 and corrected for cell viability. The results were expressed as the ratio between the percentage of infection and the percentage of viable cells. This ratio was normalized to 100% in the control samples in which only DMSO was added.