FIG. 3.

(A) Infection of ECs with KSHV had no effect on EC proliferation as determined by BrdU incorporation (n.s., not significant). Similarly, transduction of ECs with the empty LZRS vector or with LZRS-vFLIP had no effect on proliferation. In contrast, expression of v-cyclin or LANA in ECs resulted in a small, but statistically significant, increase in cell proliferation (absorbance values of 1.14 ± 0.03 for v-cyclin-transduced ECs [EC+v-cyclin] and 1.17 ± 0.04 for LANA-transfected ECs [EC+LANA] were significantly different [P < 0.05] from the 0.96 ± 0.03 value for ECs). Four independent experi-ments were performed. (B) Infection with KSHV had little to no effect on the proliferative capacity of ECs. Population doubling (PD) was calculated in three independent experiments, and a representative experiment is shown. It should be noted, however, that in several experiments, including some in which PD was not calculated, the KSHV-infected ECs (EC+KSHV) appeared to grow slower than the uninfected ECs during the early passages immediately after infection. (C) Western blot analysis of CDKI expression in KSHV-infected ECs. A representative experiment is shown where expression of the following proteins in KSHV-infected ECs (EC+KSHV) was analyzed and compared to expression in ECs: Id-1 (24.8-fold increase), p16 (14.5-fold increase), p21 (2.7-fold increase), and p27 (1.0-fold increase). The blot was probed for expression of the proteins and reprobed for actin to demonstrate equal loading of the proteins. Three independent experiments were performed and gave similar results.