Fig. 5. DYRK2 reduces sensitivity to proteotoxic stress via HSF1.

A Equal number of MDA-MB-468 WT (Left panel) or DYRK2-KO (Right panel) were seeded. After 24 h, cells were exposed to 45 °C (HS) for 45 min followed by recovery at 37 °C (cells labelled as 45 °C) or they were left at 37 °C (cells labelled as 37 °C). Number of cells at each point were analysed using the Alamar Blue assay. Data represent means ± SD (n = 3). B Control (WT) and DYRK2-KO MDA-MB-468 cells were incubated at 37 °C (−) or at 45 °C for the indicated times, followed by recovery at 37 °C. On the next day, cells were lysed and the levels of apoptosis were analysed by western blotting using an antibody that recognises cleaved PARP. The corresponding quantifications of PARP protein levels are shown in Supplementary Fig. S5A. C 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 45 °C for 45 min followed by recovery at 37 °C. On the next day, cells were lysed and the levels of apoptosis were analysed by western blotting using an antibody that recognises cleaved PARP. The corresponding quantifications of PARP protein levels are shown in Supplementary Fig. S5B. MDA-MB-468 (D) or HSF1-KO MDA-MB-468 (E) cells transfected with either siControl or siDYRK2 were incubated at 37 °C or at 45 °C for the indicated times, followed by recovery at 37 °C. On the next day, cells were lysed and the levels of apoptosis were analysed by western blotting using an antibody that recognises cleaved parp. The corresponding quantifications of PARP protein levels are shown in Supplementary Fig. S5C, D. See also Fig. S5.