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. 2020 Dec 2;28(5):1563–1578. doi: 10.1038/s41418-020-00686-8

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© The Author(s) 2020

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. 4 — A Control (WT), DYRK2-KO or HSF1-KO MDA-MB-468 cells were incubated at 37 or 42 °C for 1 h. The mRNA levels for HSP70 (HSPA1A) were quantified using real-time PCR. The data were normalised using β-actin as an internal control. Data represent means ± SD (n = 3) and are expressed relative to the control sample levels at 37 °C. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, B Control (WT), DYRK2-KO or HSF1-KO MDA-MB-468 cells were incubated at 37 or at 42 °C for 1 h followed by another hour at 37 °C. Cells were lysed and the levels of the indicated proteins were analysed by western blot. The corresponding quantifications of HSP70 protein levels are shown in Supplementary Fig. S4B. C Control (WT) and DYRK2-KO MDA-MB-468 cells were incubated at 37 °C, at 42 °C for 5, 15, 30 or 60 min, or at 42 °C for 60 min plus recovery time at 37 °C (up to 180 min). After that, mRNA levels for HSP70 (HSPA1A) were quantified using real-time PCR. The data were normalised using β-actin as an internal control. Data represent means ± SD (n = 3) and are expressed relative to the control samples at 37 °C. D DYRK2-KO MDA-MB-468 infected with virus encoding for either empty vector (Ctrl), DYRK2-WT (R. WT) or the DYRK2 HSF1-interaction deficient mutant (R. BR1 + 2) were incubated at 37 °C or at 42 °C for 60 min. After that, mRNA levels for HSP70 (HSPA1A) were quantified using real-time PCR. The data were normalised using β-actin as an internal control. Data represent means ± SD (n = 3) and are expressed relative to the control samples at 37 °C. *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001. E DYRK2-KO MDA-MB-468 infected with virus encoding for either empty vector (Ctrl), DYRK2-WT (R. WT) or the DYRK2 HSF1-interaction deficient mutant (R. BR1 + 2) were incubated at 37 °C or at 42 °C for either 60 or 120 min. Cells were lysed and the levels of the indicated proteins were analysed by western blot. The corresponding quantifications of HSP70 protein levels are shown in Supplementary Fig. S4D. See also Fig. S4.