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. 2022 Jul 19;6:52. doi: 10.1038/s41698-022-00296-2

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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. 3 — a Western blot analysis showing increase in AKT protein phosphorylation upon transfection with constitutively active AKT mammalian vector. b, c Cell viability assays showing the effect of single treatment of mutant and parental H1581 cell lines with AZD4547 for 96 h (b) or AZD4547 combined with AZD5363 after 96 h (c). d The table shows numbers and percentages of single cells with induced resistance after continuous treatment with AZD4547 in three lung-cancer cell lines. e Western blot analysis comparing FGFR1 and AKT expression and activation among the three induced-resistance single clones H1581-C4R, H1581-C9R and H1581-C11R with their sensitive parental control NCI-H1581 cell line. f Cell viability assay measuring the effect of AZD4547 treatment on the three named resistant clones. g Counting of viable cells using cell-cycler under treatment of AZD4547 and/or AZD5363. h–j Cell viability assay for combined AZD4547 and AZD5363 treatments after 96 h in sensitive parental control cell line H1581 and three FGFR1 inhibition-resistant clones H1581-C4R, H1581-C9R and H1581-C11R. k Combination index plot calculated with CompuSyn according to the Chou–Talalay equation algorithm: CI < 1, synergetic effect; CI = 1, additive effect; CI > 1, antagonistic effect. Statistical analysis was performed with the chi-squared test: ns (p > 0.05), *(p < 0.05), **(p ≤ 0.01) and ***(p ≤ 0.001). Mean values are plotted; error bars represent standard deviation.