PMC:6365940 / 43731-48001 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/6365940","sourcedb":"PMC","sourceid":"6365940","source_url":"https://www.ncbi.nlm.nih.gov/pmc/6365940","text":"Appendix A Supplementary material\nSupplementary Fig. S1 Protein expressions of ABCC1 and ABCC5 in SW1990, SW1990/Gem, Panc-1 and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P \u003c 0.001 vs. SW1990 or Panc-1 cells.\nSupplementary Fig. S2 Digoxin could not enhance the cytotoxicity of gemcitabine and inhibit the Nrf2 signaling in SW1990 and Panc-1 cells. (A–B) Effects of digoxin on chemosensitivity of SW1990 and Panc-1 cells to gemcitabine. (C–D) Effects of digoxin on the protein levels of Nrf2 in SW1990 and Panc-1 cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments.\nSupplementary Fig. S3 Digoxin enhanced the sensitivity of SW1990/Gem and Panc-1/Gem cells to etoposide, paclitaxel, cisplatin, 5-FU, ara-C and doxorubicin. Data were expressed as mean ± SD, and the results were representative of three independent experiments.\nSupplementary Fig. S4 Digoxin inhibited the protein expression of ABCC5 in SW1990/Gem and Panc-1/Gem cells and in SW1990/Gem cells-derived xenografts. (A–B) Effect of digoxin on protein levels of ABCC5 in SW1990/Gem and Panc-1/Gem cells. (C–D) SW1990/Gem and Panc-1/Gem cells were treated with 80 nM of digoxin, 20 µM of LY294002, or a combination of digoxin and LY294002 for 24 h, the protein levels of Nrf2 and ABCC5 were detected by Western blot. (E-F) Effects of Nrf2 knockdown on protein levels of Nrf2 and ABCC5 in SW1990/Gem and Panc-1/Gem cells. (G) Effects of digoxin on the protein levels of Nrf2 and ABCC5 in tumor tissues. (H) Tumor tissues were subjected to IHC-ABCC5 staining. All images were shown at ×200. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P \u003c 0.001 vs. control group or vehicle group. N.S., no significant.\nSupplementary Fig. S5 Digoxin inhibited Nrf2 not through affecting stability of Nrf2 mRNA and translation and degradation of Nrf2 protein in SW1990/Gem and Panc-1/Gem cells. (A–B) 80 nM of digoxin was incubated with or without actinomycin D (5 μg/mL) in SW1990/Gem and Panc-1/Gem cells for 2 h, 4 h and 8 h. Nrf2 mRNA levels at different time points (2 h, 4 h and 8 h) were detected using qRT-PCR. (C–D) 80 nM of digoxin was incubated with or without actinomycin D (5 μg/mL) in SW1990/Gem and Panc-1/Gem cells for 2 h, 4 h and 8 h. Nrf2 protein levels at different time points (2 h, 4 h and 8 h) were detected using Western blot. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as **P \u003c 0.01, ***P \u003c 0.001 vs. control group. N.S., no significant.\nSupplementary Fig. S6 Effect of digoxin on PI3K kinase activity. PI3K was reacted with PI(4,5)P2 in the presence and absence of digoxin for 2 h and PI(3,4,5)P3 production was measured with a competitive ELISA. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P \u003c 0.001 vs. control group.\nSupplementary Fig. S7 (A–B) Effects of Nrf2 overexpression plasmid on protein levels of Nrf2, NQO1, HO-1, GCLC and ABCC5 in digoxin-treated SW1990/Gem and Panc-1/Gem cells. (C–D) Nrf2 overexpression plasmid could restore drug resistance of gemcitabine in digoxin-treated SW1990/Gem and Panc-1/Gem cells. Data were expressed as mean ± SD, and the results were representative of three independent experiments. Significant differences were indicated as ***P \u003c 0.001 vs. control group.\n\nAcknowledgements\nThis work was supported by the National Natural Science Foundation of China (Nos. 81372268, 81672816 and 81872337), the Program for Jiangsu Province Innovative Research (KYLX16_1120), the Natural Science Foundation for Distinguished Young Scholars of Jiangsu Province (No. BK20130026), the Project Program of State Key Laboratory of Natural Medicines, China Pharmaceutical University (No. ZJ11173).\n\nConflict of interest\nThere is no conflict of interest. Appendix A Supplementary data associated with this article can be found in the online version at doi:10.1016/j.redox.2019.101131.","divisions":[{"label":"sec","span":{"begin":0,"end":3666}},{"label":"label","span":{"begin":0,"end":10}},{"label":"title","span":{"begin":12,"end":34}},{"label":"p","span":{"begin":35,"end":335}},{"label":"figure","span":{"begin":35,"end":335}},{"label":"label","span":{"begin":35,"end":56}},{"label":"caption","span":{"begin":58,"end":335}},{"label":"p","span":{"begin":58,"end":335}},{"label":"p","span":{"begin":336,"end":750}},{"label":"figure","span":{"begin":336,"end":750}},{"label":"label","span":{"begin":336,"end":357}},{"label":"caption","span":{"begin":359,"end":750}},{"label":"p","span":{"begin":359,"end":750}},{"label":"p","span":{"begin":751,"end":1011}},{"label":"figure","span":{"begin":751,"end":1011}},{"label":"label","span":{"begin":751,"end":772}},{"label":"caption","span":{"begin":774,"end":1011}},{"label":"p","span":{"begin":774,"end":1011}},{"label":"p","span":{"begin":1012,"end":1951}},{"label":"figure","span":{"begin":1012,"end":1951}},{"label":"label","span":{"begin":1012,"end":1033}},{"label":"caption","span":{"begin":1035,"end":1951}},{"label":"p","span":{"begin":1035,"end":1951}},{"label":"p","span":{"begin":1952,"end":2794}},{"label":"figure","span":{"begin":1952,"end":2794}},{"label":"label","span":{"begin":1952,"end":1973}},{"label":"caption","span":{"begin":1975,"end":2794}},{"label":"p","span":{"begin":1975,"end":2794}},{"label":"p","span":{"begin":2795,"end":3183}},{"label":"figure","span":{"begin":2795,"end":3183}},{"label":"label","span":{"begin":2795,"end":2816}},{"label":"caption","span":{"begin":2818,"end":3183}},{"label":"p","span":{"begin":2818,"end":3183}},{"label":"p","span":{"begin":3184,"end":3666}},{"label":"figure","span":{"begin":3184,"end":3666}},{"label":"label","span":{"begin":3184,"end":3205}},{"label":"caption","span":{"begin":3207,"end":3666}},{"label":"p","span":{"begin":3207,"end":3666}},{"label":"ack","span":{"begin":3668,"end":4083}},{"label":"title","span":{"begin":3668,"end":3684}},{"label":"p","span":{"begin":3685,"end":4083}},{"label":"ack","span":{"begin":4085,"end":4139}},{"label":"sec","span":{"begin":4085,"end":4139}},{"label":"title","span":{"begin":4085,"end":4105}},{"label":"p","span":{"begin":4106,"end":4139}},{"label":"label","span":{"begin":4140,"end":4150}}],"tracks":[]}