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Rhodamine 123 Accumulation Assay

YZ Yanling Zhao
HY Han Yan
XL Xue Liang
ZZ Zhenyu Zhang
XW Xuan Wang
NS Nianwei Shi
WB Weihong Bian
QD Qing Di
HH He Huang
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After drug treatment, cells were washed with PBS. After a 90 min incubation with Rhodamine 123 (2 μM) (C2007; Beyotime Biotechnology) at 37 ℃, Rhodamine 123 was washed with cold PBS and 1% Triton X-100, and the cells were kept on ice until analysis. After being incubated with the DAPI staining solution for 10 min, the accumulation of Rhodamine 123 was measured using a Leica fluorescence microscope (Leica Microsystems GmbH, Germany). The stronger the efflux activity of P-gp in cells, the weaker the fluorescence intensity of intracellular Rhodamine 123. The mean fluorescence of Rhodamine 123 in cells was calculated using ImageJ 1.46 software.

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