Figure 9.

Effect of EBNA5 on the expression of cellular genes on the chromosomes. DG75 cells were transfected with the pE300CY6 plasmid and pCI-EBNA5 or pCI. The transfected cells were purified on the basis of rCD2 expression and the poly(A)⁺ RNA fraction isolated and used as template for the preparation of ³²P-labelled cDNA. Human Atlas DNA microarray membranes (Clontech) containing cDNA fragments representing 588 human genes were incubated under hybridisation conditions with probes derived from pCI-transfected cells (upper) and pCI-EBNA5-transfected cells (lower), respectively. The figure shows the representative results of one of three independent experiments. The positions corresponding to the BNIP3, CD27L, GAPDH and β-actin cDNAs are indicated. The expression level of each gene as reflected by the amount of ³²P hybridised to the membrane was quantified by phosphorimaging. The influence of EBNA5 on expression of the genes represented on the membrane was calculated as the ratio between the hybridisation signal obtained with the probe derived from cells transfected with the EBNA5 expression vector and the probe from cells transfected with the control plasmid.