Skip to main content
. 2011 Jan 5;85(6):2666–2685. doi: 10.1128/JVI.01532-10

FIG. 5.

FIG. 5.

Effect of neomycin treatment on TIVE-LTC cell survival and viral gene expression. (A) TIVE-LTC cells were treated with increasing concentrations of neomycin for 3 days and assayed for cell survival by MTT assay. Paromomycin (100 μM) treatment was used as a control. (B) The trypan blue assay was done with TIVE and TIVE-LTC cells at 0, 1, 2, and 3 days posttreatment with 200 μM neomycin, and the results are represented as the percentage of cell survival by considering the number of cells in untreated cells on each day as 100%. (C and D) cDNA prepared from TIVE-LTC cells left untreated (UT) or treated with 200 μM neomycin after 24, 36, and 48 h was used for measurement of ORF73 (C) and ORF50 (D) gene expression by quantitative real-time PCR and normalized to expression of tubulin. (E) TIVE-LTC cells treated with the indicated concentrations of neomycin for 3 days were stained with antibodies against ORF73, ORF59, and gpK8.1A. Cells were washed with PBS, incubated with secondary antibody conjugated with Alexa Fluor 488 (green) for 1 h at room temperature, washed, mounted, and visualized under a fluorescence microscope. Magnification, ×20. (F) TIVE-LTC cells were left untreated or treated with 200 μM neomycin for 3 days, virus was purified from the supernatant, DNA was extracted, and ORF73 DNA copy numbers were determined. **, P < 0.001; ***, P < 0.0001.