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The osmotic pressure difference required to open MscL. The G22C mutant of E. coli MscL was reconstituted into vesicles of a 1:1 mixture of DOPA:DOPC containing 50 mM calcein and diluted into buffer containing 100 mM KCl and the following concentrations of sucrose: (a) 0.5 M; (b) 0.45 M; (c) 0.4 M; (d) 0.2 M; (e) 0 M. After 150–250 s, 200 μM C12E8 was added to burst the vesicles.
