Fig. 7. PIDDosome deficiency reverses PLK4-induced delays in IR-driven tumor onset.

(A) Kaplan-Meier analysis of tumor-free survival. rtTA (n = 20), median survival 143.5 days; rtTA/Plk4 (n = 16), median survival 174.5 days. Log-rank (Mantel-Cox) P = 0.0328 (χ² = 4.554), Breslow-Gehan-Wilcoxon P = 0.0232 (χ² = 5.157). rtTA/Plk4 (n = 16) versus rtTA/Plk4/Raidd^−/− (n = 15), median survival 174.5 days; rtTA/Plk4/Raidd^−/−, median survival 158 days. Log-rank (Mantel-Cox) P = 0.2104 (χ² = 1.569), Breslow-Gehan-Wilcoxon P = 0.0644 (χ² = 3.419). (B) Tumors were isolated from mice fed with doxycycline-containing food after IR and were analyzed for centrosome amplification by IF using antibodies recognizing CP110 and γ-tubulin (two biological and two technical replicates). (C) Quantification of Western blots for p53 and p19ARF to estimate p53 function in respective tumors isolated from terminally ill mice with the indicated genotypes. (D) Kaplan-Meier analysis of tumor-free survival of rtTA transgenic mice that were defined to have developed p53 wild-type (wt) (n = 16) or null (n = 6) tumors (median survival 156 versus 131.5 days). Log-rank (Mantel-Cox) P = 0.2128 (χ² = 1.552), Breslow-Gehan-Wilcoxon P = 0.0332 (χ² = 4.537). (E) Tumor-free survival of rtTA/Plk4 transgenic mice that were defined to have developed p53 wt (n = 11) or null (n = 12) tumors (median survival 179 versus 174 days). Log-rank (Mantel-Cox) P = 0.8194 (χ² = 0.05212), Breslow-Gehan-Wilcoxon P = 0.9756 (χ² = 0.0009). (F) Tumor-free survival of mice defined as having developed p53 null tumors on an rtTA (n = 6) or rtTA/Plk4 background (n = 12), median survival 131.5 versus 174 days. Log-rank (Mantel-Cox) P = 0.0339 (χ² = 4.5), Breslow-Gehan-Wilcoxon P = 0.0118 (χ² = 6.337). *P < 0.05, **P < 0.01, ***P < 0.005. (G) Tumor cell lines generated from rtTA (four biological replicates), rtTA/Plk4 (n = 1), rtTA/p53mut/del (n = 1), and rtTA/Plk4/p53mut/del (two biological replicates) mice were treated with doxycycline (1 μg/ml) for 48 hours or 1 μM etoposide for 12 hours to induce cell death measured by annexin V/DAPI staining. Up to three technical replicates were performed.