Fig. 4

Mutp53 is associated with elevated mRNA and protein levels of NEK2. a Heat map showing cluster analysis of differentially expressed genes in p53+/+;ErbB2, p53H/+;ErbB2 and p53−/+;ErbB2 before and after γ-irradiation (9 Gy, 24 h). b Heat map of RNAseq analysis showing differentially regulated centrosome proteins in p53+/+;ErbB2, p53H/+;ErbB2 and p53−/+;ErbB2 before and after γ-irradiation (9 Gy, 24 h). Arrowhead indicates Nek2. c Western blot analysis of Nek2 level before and 24 h after γ-irradiation in mouse ErbB2 mammary epithelial tumor cell lines with different p53 status. α-tubulin is loading control. d Bar graph showing relative mRNA expression level of Nek2 in p53H/−;ErbB2 and p53−/−;ErbB2 cell lines before and after irradiation. n = 3 independent experiments per genotype (one cell line per genotype except for p53 H/− where 2 different cell lines derived from different tumors and result was averaged). e Western blot analysis of p53 and Nek2 levels in p53−/−;ErbB2 cells and in p53H/−;ErbB2 cells before and after CRISPR/Cas9 p53 deletion. Hsp70 is loading control. f Western blot analysis of Nek2 level in p53−/− MECs and p53H/H MECs. α-Tubulin is loading control. g Western blot analysis of p53 and Nek2 levels in BT474 cells before and after p53 suppression with siRNA. HSC70 is loading control. h Bar graphs showing NEK2 relative mRNA expression in patients with wtp53 (n = 1245) compared to with mutp53 (n = 659) (all mutations combined vs different types of mutations) (DBD = DNA binding domain; TD = tetramerization domain). i Bar graph showing Nek2 relative mRNA expression in human breast cancer cell lines with different p53 status. n = 3 independent experiments per genotype. Where applicable *p < 0.05; **p < 0.01; ***p < 0.001. Error bars represent ± SD