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. 2020 Mar 6;12(3):608. doi: 10.3390/cancers12030608

Figure 4.

Figure 4

Impact of drug hits on expression of copper transporters and Pt-mediated trafficking of ATP7B in IGROV-CP20 cells. (A–F) IGROV-CP20 cells were treated with 10 µM Tranilast, Telmisartan, or Amphotericin B for 24 h and then 50 µM cisplatin was added for 24 h. The cells were then prepared for qRT-PCR (A,C,E) or Western blot ((B,D,F); see also signal density quantification in each panel) to evaluate the expression of ATP7B (A,B), ATP7A (C,D), and CTR1 (E,F) (for each qRT-PCR and blot n = 3 experiments; * p < 0.05, ** p < 0.01, ANOVA). (G,H) The cells were treated with 10 µM Tranilast, Telmisartan, or Amphotericin B for 24 h and then exposed to 50 µM cisplatin for 4 h to activate ATP7B trafficking. Confocal images (G) show that the drugs promote ATP7B retention in the Golgi (labeled with Golgin-97) upon cisplatin treatment as also revealed by quantification of the ATP7B signal (H) in the Golgi region (n = 10 fields; ** p < 0.01, *** p < 0.001, ANOVA). Scale bar: 15 µm (G).