The gammaherpesvirus 68 viral cyclin facilitates expression of LANA
Fig 3
Disruption of the viral cyclin has no effect on the frequency of viral genome positive cells yet results in a reduced frequency of LANA::βla+ cells.
Mice were infected via IP injection with WT.βla or cycKO.βla viruses and splenocytes were harvested at 8 or 16 dpi. (A) Limiting-dilution nested PCR of viral gene ORF 50 from WT.βla or cycKO.βla infected splenocytes. n = 4 (8 dpi) or n = 3 (16 dpi) with 3–5 mice pooled per group with SEM shown. Comparisons between the LogEC(63.2) found statistical difference between the of WT.βla and cycKO.βla at 8 dpi only (p = 0.009). (B) Representative pseudocolor plots identifying LANA::βla+ splenocytes, indicated in the upper right polygon. Average frequencies of LANA::βla+ cells +/- SEM is indicated below the gate. 8 dpi is shown on the left, while 16 dpi is shown on the right, with WT.βla infected mice on top and cycKO.βla infected mice on bottom. (C) Percent of LANA::βla+ cells (top) and total number of LANA::βla+ splenocytes (bottom) from each individual mouse plotted with SEM shown after infection with WT.βla (black) or cycKO.βla (red) virus. (D) Graphical representation of the number of LANA::βla+ cells expressing cell surface markers with SEM shown after infection with WT.βla (black) or cycK.βla (red) virus. Cells were stained and gated as in Figure 2.8 dpi: LANA::βla+, CD19, IgD, and CD38 n = 13 and CD44 n = 7.16 dpi n = 11. Two-tailed student t tests were performed to measure statistical significance in C and D.