3 sn pa

Powder form of 3-sn-PA (Sigma Aldrich), Brain PS (Avanti Polar Lipids), Brain PI-(4)-P (Avanti Polar Lipids) was dissolved in chloroform in a clean glass vial and evaporated in a desiccator overnight at RT. Thin film of dried lipid obtained after drying was dissolved in 20 mM Tris pH 7.5 and 150 mM NaCl and stored in −20 °C for future use.

Powder form of Egg-PC (Sigma Aldrich), L-α-phosphatidylethanolamine (Sigma Aldrich), 16:0 Cardiolipin (Avanti Polar Lipids), 3-sn-PA (Sigma Aldrich), Brain PS (Avanti Polar Lipids) was dissolved in chloroform. Next, 270 μM PC, 145 μM phosphatidylethanolamine, 65 μM PIP4, 5 μM CL, 5 μM PA, 10 μM PS were mixed and kept for overnight evaporation. The thin lipid film obtained the following day after removal of residual chloroform was dissolved in 500 μl of 20 mM Tris pH 7.5 and 150 mM NaCl with gentle pipetting. The solution was next incubated at 55 °C for 30 min. The resultant colloidal heated solution was subjected to extrusion as mentioned above. Finally, the model membrane liposomes comprising a mixture of PC, phosphatidylethanolamine, PIP4, CL, PA, and PS were used for downstream experiments and the remaining was stored at 4 °C for not more than 3 days.