p27 expression in B-cells infected with Δ123.

<p>(a) We performed a BrdU incorporation assay with LCLs transformed by wild type or Δ123 viruses. The figure shows the results of the FACS analysis two months after infection. (b) 2 months-old cells infected with wild type or the Δ123 virus were brought to a concentration of 3x10⁵ cells per ml and the increase in cell numbers was determined by counting trypan blue-negative cells 24 and 48 hours thereafter. The growth rate is given relative to initial cell numbers. The growth kinetics of four donors were determined twice in independent experiments. (c) Western blot for PTEN expression from one representative donor infected with B95-8 wt or B95-8 Δ123. LCLs were analyzed 2.5 months after their establishment. Actin served as a loading control. ImageJ quantified PTEN expression in three donors 2.5 month after infection is shown in the right panel. All values were normalized for differences in actin expression. (d) We measured p27 expression in seven 2–3 months old LCLs established with wild type EBV or with Δ123 using a western blot analysis. The left panel shows one example; the right panel gives the results of ImageJ-based quantification for all tested donors. The individual values were normalized for differences in actin levels and p27 levels are depicted relative to the respective wild type control. (e) p27 expression was monitored between day 1 and 18 after infection of primary B-cells with wild type or Δ123 viruses. Actin served as a loading control. The right panel depicts the ImageJ-based quantification of the p27 protein levels normalized for differences in actin expression.