A sine wave of frequency 10 Hz was used for electroformation at different temperatures depending on the membrane composition with the peak to peak voltage changing every 10 minutes from 0.1 to 0.5, 1, 2 and finally 3 V for 2 hours. 0-50 mol% PBd12-PEO11, electroformation was carried out at 35 °C, 25 and 50 mol% PBd22-PEO14 at 42 °C, and 75 and 100 mol% of both polymers at 64 °C. The frequency was gradually decreased over approximately 8 min to facilitate the closure and detachment of GUVs from the surface. After electroformation, the GUVs were suspended in an isoosmolar 20 mM HEPES, 150 mM NaCl solution (confirmed by measurement using the micro-osmometer).