Journal: bioRxiv
Article Title: Potent synthetic lethality between PLK1 and EYA-family inhibitors in tumours of the central and peripheral nervous system
doi: 10.1101/2025.01.19.633804
Figure Lengend Snippet: (A-B) Quantification of mean EYA3 or EYA2 protein levels relative to IMR32 via densitometry from (two-tailed t test, ns = non-significant). (C) IMR32 cell CETSA assay western blots of EYA1-4 following treatment with DMSO or Benzarone (10 µM, 2 hrs) and exposure of lysates to the indicated temperatures. (D) Densitometry quantitation of the CETSA assay. EYA1-4 band densitometry values are expressed relative to the respective value in the 37°C treatment. Greater values for each EYA at increasing temperatures in the benzarone treated samples compared to DMSO indicates target binding. (E) Relative viability across combination dose-response matrixes (Benzarone and Volasertib) measured by alamarBlue in H4 cells (n=3, mean ZIP synergy score is shown below) (F) Relative viability across combination dose-response matrixes (Benzarone and Volasertib) measured by alamarBlue in T98G cells (n=3, mean ZIP synergy score is shown below).
Article Snippet: Western blot primary antibodies targeted EYA1, Protein Tech 22658 (1:1000), EYA2, Millipore HPA027024 (1:1000), EYA3, Protein Tech 21196 (1:1000), EYA4, Santa Cruz SC393111 (1:1000), Actin, Sigma a2066 (1:2000), PARP, Cell Signalling 9542 (1:1000), gamma-H2AX Millipore 05-636 (1:1000), Vinculin, Sigma V9131 (1:4000), p-S10 H3 Cell Signalling 9706 (1:1000), CD133, Cell Signalling 64326 (1:1000), and Sox9, Cell Signalling 82630 (1:1000).
Techniques: Two Tailed Test, Western Blot, Quantitation Assay, Binding Assay