mLANA DNA binding is essential for virus persistence and the dorsal positive patch exerts a role in the expansion of GC B cells.

<p>(<b>A</b>) Amino acid substitutions in recombinant viruses (see also <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003673#ppat.1003673.s005" target="_blank">Figure S5</a>). (<b>B</b>) Infection of BHK-21 cells at 0.01 p.f.u. per cell. Virus titres were determined by plaque assay. (<b>C</b>) Lungs from infected mice were removed and infectious viruses were titrated by plaque assay. (<b>D and E</b>) Quantification of latent infection in spleen by explant co-culture plaque assay (closed circles). Titres of infectious virus were determined in freeze/thawed splenocyte suspensions (open circles). Each circle represents the titre of an individual mouse. The dashed line represents the limit of detection of the assay. Mutant viruses are shown in panel D and revertant viruses in panel E. (<b>F and G</b>) Reciprocal frequencies of viral DNA-positive cells in total splenocytes (<b>F</b>) or GC B cells (CD19⁺CD95^hiGL7^hi) (<b>G</b>) were determined by limiting dilution and real-time PCR. Data were obtained from pools of five spleens per group. Bars represent the frequency of viral DNA-positive cells with 95% confidence intervals. (<b>H</b>) Identification of latently infected cells in spleens by <i>in situ</i> hybridization. Representative splenic sections from each group of viruses are shown. All images are magnified ×200. Dark staining indicates cells positive for virally encoded miRNAs (see also <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.1003673#ppat-1003673-t004" target="_blank">Table 4</a>).