Figure 2

PML aggregation occurs specifically in transfected cells. pEGFP Neo empty vector was transiently cotransfected with pBABE Puro empty vector in U2OS cells on glass coverslips. Cells were immunostained for PML (red) 48 h after transfection. PML antibodies were detected using a rhodamine-x conjugated goat anti mouse IgG. Cell nuclei were counterstained with DAPI (blue). Transfected cells were identified by EGFP expression (green). (A) Representative images of U2OS cells transfected with pEGFP Neo empty vector alone or pEGFP/pBABE Puro demonstrating PML aggregation in co-transfected cells. Images captured at 100× magnification. (B) Percentage of co-tansfected EGFP positive cells displaying PML aggregation were quantified. Percentage represents average ± SEM of five independent experiments with 300 cells counted per experiment.