CF release liposome assay.

<p>(A) aliquots of p7 in methanol (M), TFE (TFE) or HFIP (H) were added to CF-loaded PAESC liposomes and fluorescence values were measured every minute; (B) Comparison of CF release results of full length p7 versus p7 fragments and melittin. Experimental conditions were as in (A), except that each peptide was added in methanol and peptide∶lipid molar ratio was 1∶10. Error bars (1 SD) are the result of 3 independent measurements; (C) same experiment, but p7 was incorporated into the liposomes by the dyalisis method after freeze-thaw and extrusion. Fluorescence values were measured immediately (0 h) after removing excess CF and after one hour (1 h). Liposome without p7 was used as control, and TX100 was added to measure total amount of CF incorporated. Error bars (1 SD) are shown only for one trace for clarity, and are the result of 3 independent measurements; In (A) and (B), no effect was observed when the liposomes were preincubated with up to 100 µM rimantadine; (D) kinetics of CF release when liposomes were extruded, where p7(1) and p7(2) represent samples containing p7 reconstituted with or without detergent solubilization steps, respectively.