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. 2017 May 17;51(1):257–268. doi: 10.3892/ijo.2017.4005

Figure 5.

Figure 5

Icory stimulates ABCB1 ATPase activity but does not alter ABCB1 expression or localization; the MDR-reversal effect of Icory is reversible. (A) The effect of Icory at 25 and 50 μM on the mRNA expression of ABCB1 in HepG2/ADM and MCF-7/ADR cells at 24 and 48 h; GAPDH was used as an internal control. (B) Effect of Icory at 25 and 50 μM on the protein expression of ABCB1 in HepG2/ADM and MCF-7/ADR cells at 24 and 48 h; β-actin was used as an internal control. (C) Effect of Icory at 50 μM on the localization of ABCB1 in HepG2/ADM cells for 24 and 48 h. ABCB1 staining are shown in green. DAPI (blue) counterstains the nuclei. Original amplification, 630; bar, 10 μm. (D) The effect of Icory on ABCB1 ATPase activity was detected using the Pgp-Glo™ ATPase assay kit. VRP was used as a positive-control inhibitor of ABCB1. (E) Icory had no continued MDR-reversal effect after removal. The cells were pre-incubated with or without Icory (50 μM) or VRP (50 μM) for 24 h; the compound was then washed out, and the cells were incubated with DOX for 72 h. The viability of the cells was tested by the MTT assay. ***P<0.001 compared with the Icory-washout group and ###P<0.001 compared with the VRP-washout group. Each point represents the mean ± SD of at least three independent experiments performed in triplicate. *P<0.05, **P<0.01 and ***P<0.001 compared with the control.