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. 2009 Jan;11(1):96–101. doi: 10.1593/neo.81264

Figure 3.

Figure 3

(A) Fluorescent dye uptake assay indicates that HhAntag691 inhibits Pgp and MRP1 but not MRP2. MDCKII cells overexpressing Pgp, MRP1, or MRP2 were incubated in medium containing no drug or different ABC transporter inhibitors (50 µM VP, 50 µM indomethacin, or 20 µM HhAntag691), then calcein-AM was added to a final concentration of 1.0 µM, and incubated at 37°C for 1 hour. Cells were then washed and lysed, and calcein fluorescence was determined (excitation wavelength, 495 nm; emission wavelength, 515 nm). All readings are normalized to control when no drug was added. (B) HhAntag691 causes a dose-dependent increase of fluorescent signal retention of calcein-AM in MDCKII/Pgp cells. Cells were incubated at 37°C in medium containing 1 µM calcein-AM and increasing concentrations of HhAntag691 for 2 hours and were then washed and lysed, and fluorescent signal was quantified (excitation wavelength, 495 nm; emission wavelength, 520 nm). Mean ± SEM, n = 3.