HBMEC, a cell model for BBB, was established on the upper chamber of a Transwell insert. C6-PLs or C6-cRGDLs (equivalent to 1 μM SPC) with M/Ns (1 × 105 cells) or without M/Ns were added onto the HBMEC monolayer, which had been pretreated with 10−7 M fMLP in the lower chamber. After 1 hour of incubation, the suspension in the lower chamber was collected and subjected to an ultrasonic treatment for C6 fluorescence assay using the microplate reader. Blank M/Ns (not treated with liposomes) were used as blank controls for cells, and non–fluorescence-labeled liposomes (PLs and cRGDLs) were used as a blank control for liposomes (Fig. 1D). The TEER (trans epithellal electric resistance) of the in vitro barrier model was monitored using a resistance instrument during the experiment (data are shown in fig. S9A).

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