Drug distribution analysis of the drug-loaded cells
This protocol is extracted from research article:
Transport of a graphene nanosheet sandwiched inside cell membranes
Sci Adv, Jun 7, 2019; DOI: 10.1126/sciadv.aaw3192

Cells were seeded in a 10-cm plate overnight and were exposed to different drug solutions. After cell-drug interaction for 24 hours, 107 cells were harvested and washed twice with PBS. To determine the drug distribution amount, cell membrane fractions were isolated by using a Membrane and Cytosol Extraction Kit. The drugs from different cell fractions were then extracted from the water phase (cell fraction suspensions) to the oil phase (chloroform). The amounts of drug importing into the membrane and total drug entering the cell were finally detected by using a liquid chromatography mass spectrometer (Thermo Scientific TXQ Quantum ACCESS Max) and by using a selective reaction monitoring model. To further delineate the relationship between the sandwich structure and the intracellular drug amount, the mean fluorescence intensity of cells was assayed by flow cytometry (Cytoflex), where the fluorescence was sourced from the sandwiched GO upon excitation by an 808-nm NIR laser.

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