In order to evaluate the role of initial hydration and ionization level of the LNP, we modified the structure of LNP with Lipid 2. We reduced the number of water beads up to a full dehydration of the initial structure (90, 80, 50 and 0% of original water level). Further, we reduced the number of Cl -ions to 90, 50 and 0% of original level at the fully hydrated structure and to 50 and 0% at the fully and 50 % dehydrated LNP structure (Figure ). The LNPs were simulated for 10 𝜇s with annealing as other LNPs in this paper and their level of their hydration was evaluated as in ref .