Figure 7

Expression of EGFP is dependent on de novo synthesis. To exclude the possibility that the increase in fluorescence after virosome or immunovirosome (containing anti-CD71 mAb) gene transfer was due to transfer of fluorescent protein by viruses, irradiated viruses were used (A). Alternatively, ECs were treated with cycloheximide to prevent de novo synthesis of protein (B). Furthermore, to exclude that the increase in fluorescence was due to the carryover transfection of EGFP plasmids, the supernatants were treated with DNAse (C). Unstimulated HSVECs were treated with MMLV-EGFP + polybrene, virosomes, or immunovirosomes. The numbers of EGFP-expressing cells were analyzed by flow cytometry on day 3. All results are expressed as mean ± SD of triplicate wells and all experiments were carried out at least three times.