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. 2007 Jun 8;3(6):e80. doi: 10.1371/journal.ppat.0030080

Figure 3. Induction of EBV Lytic Cycle in Akata-GFP Cells upon Stimulation with CIDR1α.

Figure 3

(A) Akata-GFP cells were cultured in medium alone, with GST (4 μM), with CIDR1α (0,12–4 μM), or with anti-Ig (10 μg/mL). After 48 h of incubation, a FACS analysis was performed. Cell populations were divided into cells with low granularity (LG) and cells with high granularity (HG) on the basis of side scatter (SSC). The proportion of cells in lytic cycle (GFP-positive) was then quantified in the HG subpopulation. The figure shows one representative experiment. FSC, forward scatter.

(B) Akata-GFP cells were cultured with GST (2 μM), with CIDR1α (0–2 μM), and with anti-Ig (10 μg/mL). After 48 h of incubation, the proportion of GFP-positive cells was quantified by FACS analysis (circles), and the EBV DNA load was assessed by real-time PCR (diamonds). The left y-axis denotes the proportion of Akata-GFP–positive cells in lytic cycle, the right y-axis denotes the number of EBV genomes. Controls included GST-stimulated (empty diamonds/circles) and anti-Ig–stimulated cells (filled diamonds/circles). Data show the mean of three independent experiments.

(C) Akata cells up-regulate BZLF1 upon stimulation with CIDR1α. Akata-GFP cells were cultured with GST (1,5 μM) or with CIDR1α (0,25–1,5 μM). After 48 h of incubation, the expression of BZLF1 was analyzed by Western blot. Increased BZLF1 expression in CIDR1α stimulated samples was confirmed by relative quantitative analysis with Quantity One software (Bio-Rad).