The Ad/Liposome complex shows enhanced transfection capacity in B16-F10 cells. B16-F10 cells were transfected at different liposome and Ad-GFP ratio. After infected for 48 h, the percentage of GFP expression was determined by FACS analysis (a). Three replicates were performed for each treatment. The data shown are mean ± s.d. (*P < 0.05). Expression of PEDF protein in B16-F10 cells was determined by western blot (b). Lane1: transfected with Ad-PEDF liposome complex (Ad-PEDF/Liposome) at a ratio of 5 × 107 IFU/10 μg; lane2: transfected with Ad-PEDF alone (Ad-PEDF); Lane3: untreated. Then, B16-F10 cells were transfected with Ad-GFP (c) and Ad-GFP/Liposome (d) at a ratio of 5 × 107 IFU/10 μg. Besides, neutralizing antibodies of Ad vector pre-incubated B16-F10 cells were also transfected with Ad-GFP/Liposome complex at the same concentration (d). Ad-GFP/Liposome complex transfection noticeably enhanced the efficacy of Ad than that in Ad-GFP transfected B16-F10 cells. However, neutralizing antibodies against Ad vector only partially downregulated GFP expression (e) and data are expressed as percentages (f). Bars, SD; columns, mean.