Cholesterol contributes to membrane functionality by maintaining membrane plasticity, while simultaneously impeding the permeation of xenobiotics and ions . Its role in reducing membrane water permeability suggests that AMPs may face challenges in transporting water molecules during the initial pore-formation steps. Additionally, cholesterol promotes lateral membrane heterogeneity, leading to distinct membrane domains with varying susceptibilities to AMPs. These phenomena -membrane thickening, lipid condensing, reduced water permeability, and increased lateral heterogeneity -highlight cholesterol's particular role and provide key insights for designing potentially more selective peptides.