Percent calcein release profiles at 42C for each population. When both membranes were thermoresponsive, the fluorescence increase was much higher than when only the inner one was, because, in the latter, calcein remained encapsulated in a small enough volume to prevent unquenching. The release profiles for the additional controls mentioned in the main text can be found in Supplementary Fig. 6 , whereas compositional details can be found in Supplementary Fig. 7 . Values were calculated from fluorescence intensities after addition of surfactant (Triton X-100, 5wt%, 2.5l min –1 ). Error bars indicate the s.d. of the average intensities for n =3. The <20% calcein release for Thermo Inner: Non-thermo Outer concentrisomes was thought to originate from unilamellar vesicles that had not undergone click chemistry.
