Figure 4.

Cytosolic expression of GFP-Apoptin and quantification of apoptosis in 1BR3 normal human fibroblasts. Normal (1BR3) and transformed (1BR3LT) human fibroblast cell lines were transfected by nucleofection with pCAV-GFP-AP, pHGyV-GFP-AP or the corresponding pEGFP-C1 control plasmid as well as pCAV-FLAG-AP and pHGyV-FLAG-AP. After 3 days cells were fixed and counterstained with DAPI to analyse the subcellular localisation of GFP-Apoptin. Expression of FLAG-AP was detected using primary mouse anti-FLAG and secondary FITC anti-mouse antibody (a). Apoptosis was quantified by scoring GFP- or FLAG-positive cells containing condensed or fragmented nuclei (b). Over one hundred transfected cells were analysed and experiments were repeated twice (error bars indicate standard deviation)