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. 2020 Jan 1;11(1):25–40. doi: 10.7150/jca.32765

Figure 4.

Figure 4

The MDM2/MDMX inhibitor in combination with DOX reduces rhodamine 123 efflux in drug-resistant breast cancer cells. The cells were treated with the indicated agents for 24 h, and intracellular rhodamine 123 levels were measured to explore the transporter activity of the membrane pump protein P-gp. The concentration levels of each agent were used as follows: DOX 0.08 µg/ml in MCF-7/DOX and 0.20 µg/ml in ZR-75-30/DOX, MDM2/MDMX inhibitor 9.58 µg/ml in MCF-7/DOX and 13.70 µg/ml in ZR-75-30/DOX. The representative charts (A) and quantified data (B, C) of the rhodamine 123 assay are shown. The expression levels of the proteins investigated were examined by western blot analysis, and GAPDH was used as a loading control. The representative charts (D) and quantified data (E, F) of the western blot analysis are also shown. The values presented are indicative of the mean ± SD for each group. *P<0.05, **P<0.01, ***P<0.001 (n = 3) vs. the control group, the DOX group and/or the MDM2/MDMX inhibitor group. Immunofluorescence staining was also used in drug-resistant breast cancer cells to confirm the expression of the proteins investigated. The representative charts (G) of three independent experiments are shown.