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. 2023 May 24;13:8435. doi: 10.1038/s41598-023-35712-5

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© The Author(s) 2023

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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Co-treatment of AZ10606120 (AZ) and temozolomide (TMZ) in U251 cells. U251 human glioblastoma cells were cultured and treated at 80% confluency with various concentrations of AZ, 50 μM TMZ and a combination of AZ + TMZ for 72 h. Cells were fixed and stained with DAPI for 1 h at room temperature. The total number of DAPI + cells were quantified across 16 random fields for each sample and treatment group. (A) Comparison of AZ, TMZ and AZ + TMZ with AZ 1 μM in U251 cells. Results were yielded from a one-way ANOVA with Tukey’s HSD post-hoc, F(2, 33) = 3.77, p = 0.03. N = 12 replicates. TMZ versus AZ 1 μM: mean difference = 659.8 cells, 95% CI = − 562.5 to 1882, p = 0.39; TMZ versus AZ 1 μM + TMZ: mean difference = 1299 cells, 95% CI = 76.46 to 2521, *p = 0.04; AZ 1 μM versus AZ 1 μM + TMZ: mean difference = 639 cells, 95% CI = − 583.4 to 1861, p = 0.41. (B) Comparison of AZ, TMZ and AZ + TMZ with AZ 5 μM in U251 cells. Results were yielded from a one-way ANOVA with Tukey’s HSD post-hoc, F(2, 33) = 4.05, p = 0.03. N = 12 replicates. TMZ versus AZ 5 μM: mean difference = 958.9 cells, 95% CI = − 528.6 to 2446, p = 0.27; TMZ versus AZ 5 μM + TMZ: mean difference = 2024 cells, 95% CI = 536 to 3511, **p = 0.006; AZ 5 μM versus AZ 5 μM + TMZ: mean difference = 1065 cells, 95% CI = − 422.9 to 2552, p = 0.20. (C) Comparison of AZ, TMZ and AZ + TMZ with AZ 15 μM in U251 cells. Results were yielded from a one-way repeated measures ANOVA with Tukey’s HSD post-hoc, F(2, 30) = 3.72. N = 12 patients. TMZ versus AZ 15 μM: mean difference = 5152 cells, 95% CI = 3728 to 6576; TMZ versus AZ 15 μM + TMZ: mean difference = 6013 cells, 95% CI = 4552 to 7474; AZ 15 μM versus AZ 15 μM + TMZ: mean difference = 860.7 cells, 95% CI = − 629.9 to 2351, p = 0.34. ****p < 0.0001. (D) Comparison of AZ, TMZ and AZ + TMZ with AZ 25 μM in U251 cells. Results were yielded from a one-way repeated measures ANOVA with Tukey’s HSD post-hoc, F(2, 33) = 3.31, p = 0.049. N = 12 patients. TMZ versus AZ 25 μM: mean difference = 8848 cells, 95% CI = 7891 to 9805; TMZ versus AZ 25 μM + TMZ: mean difference = 9609 cells, 95% CI = 8652 to 10,567; AZ 25 μM versus AZ 25 μM + TMZ: mean difference = 761.4 cells, 95% CI = − 196.1 to 1719, p = 0.14; ****p < 0.0001. Error bars represent SEM.