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. 2022 Jun 13;13(6):547. doi: 10.1038/s41419-022-04983-7

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A The concentration-effect curve showed the effect of IOX1 and ATRA combination at 1:2 ratio. B Combination index value in combinatorial drug treatments. UMUC3 and 5637 cells treated with IOX1 and ATRA in combination at indicated concentrations for 48 h. Cell viabilities were measured and normalized to DMSO control values. Combination index value (CI) of each drug combination condition was calculated using CalcuSyn software. CI < 1 indicated a synergistic effect between the two drugs. C Flow cytometry analysis of early apoptosis staining by annexin V and PI and cell cycle staining by PI in UMUC3 and 5637 cells treated with IOX1 and/or ATRA. D sphere formation assay of UMUC3 and 5637 cells treated with IOX1 and/or ATRA, the sphere size was measured on day seven. E Invasion assay of UMUC3 and 5637 cells treated with IOX1 and/or ATRA. Cell Invasion was assessed by counting the number of migrating cells after 24 h. F Bioluminescence in vivo imaging showed metastasis (n = 6 per group) of nude mice injection (i.v.) of luciferase-expressing UMUC3 cells. The IOX1 and/or ATRA were treated (i.p.) daily for 10 days. IOX1, 10 mg/kg, and ATRA, 20 mg/kg. *P < 0.05; **P < 0.01; ***P < 0.001 is based on the Student’s t test. All results are from more than three independent experiments. Values are mean ± SD.