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Osmotic pressure (Π) induces membrane tension in cell membranes and the lipid bilayers of vesicles and plays an important role in the functions and physical properties of these membranes. We recently developed a method to determine quantitatively the membrane tension of giant unilamellar vesicles (GUVs) under Π and applied it to GUVs comprising electrically neutral dioleoylphosphatidylcholine (DOPC). Here, we examined the effect of Π on GUVs composed of DOPC and negatively charged dioleoylphosphatidylglycerol (DOPG) in a buffer containi