Tissue genotype, phenotype, and survival of mice with or without Pten deletion and Nf1 mutation. (A) Kaplan-Meier analysis demonstrated the high penetration (100%) of disease and early lethal effect of Pten deletion that was induced at PND8 in mice with or without Nf1 mutation. WT mice were euthanized after age 120 days. Survival in mice with PtenΔ/Δ; Nf1^+/− was significantly shorter, with a median survival of 26 days (95% confidence interval [CI], 22-32) (n = 13), and differed from PtenΔ/Δ; Nf1^+/+ (35 days [95% CI, 32-46]; P = .001), Pten^+/−; Nf1^+/−, Pten^+/+; Nf1^+/−, and WT mice (P < .001). Survival in mice with PtenΔ/Δ; Nf1^+/+ significantly differed from littermates with Pten^+/−; Nf1^+/− or Pten^+/+; Nf1^+/− and with WT (P < .001). (B) Genotype analysis by polymerase chain reaction on tissues that were collected at age 3 to 4 weeks from the littermate mice received poly(I:C) at PND8, demonstrating that Pten was deleted in the PB, spleen, and liver of mice with Pten^fl/fl and Mx1-Cre⁺, whereas the WT Nf1 allele was detectable in mice with all genotypes, suggesting that the Nf1 LOH had not occurred in diseased mice. In agreement with prior studies regarding Nf1, the intensity of signals of the mutant and WT Nf1 alleles combine to roughly equal totals. Thus, when a mutant Nf1 allele is present, the density of the WT allele is proportionately decreased (haploinsufficient). (C) Average spleen and liver weights demonstrate hepatosplenomegaly in PtenΔ/Δ mice. Data are presented as mean ± SD, *P < .05; **P < .01; ***P < .001. (D) Photographs of organs from representative littermate mice at age 4 weeks demonstrating hepatosplenomegaly; however, the thymus was spared in PtenΔ/Δ mice regardless of Nf1 background, in comparison with WT littermates (additional relevant data are presented in supplemental Figures 1 and 2). SD, standard deviation.