Hyperglycemia and pancreatic atrophy in E2F1/E2F2 compound-mutant mice. (A) Life span data obtained from WT (n = 40), DKO male (DKO M; n = 25), and DKO female (DKO F; n = 25) mice were analyzed using a log-rank nonparametric test (P < 0.01) and expressed as Kaplan-Meier survival curves. (B) Spot blood glucose levels in WT, E2F1^_/_, E2F2^_/_, and DKO mice were determined at the indicated times. Results are the means ± SEM for five to ten animals per sex and genotype. Data were analyzed using the two-tailed t test (***P < 0.001). (C) Insulin and glucagon levels in serum of 2-month-old and 6-month-old WT and DKO mice (n ≥ 7 per genotype; *P < 0.05; **P < 0.01). (D) Insulin-tolerance test was performed in starved animals following intraperitoneal injection of insulin into 5- to 6-month-old hyperglycemic female DKO mice (n = 8) or normoglycemic female WT controls (n = 7). Results are expressed as percentage of initial blood glucose concentration (*P < 0.05). Similar results were obtained for males (not shown). (E) RT-PCR analysis of RNA samples from 2-week-old WT, E2F1^_/_, E2F2^_/_, and DKO (n = 2 per genotype) mice showing expression of E2F1, E2F2, E2F5, and E2F6 in the pancreas of WT mice (E2F3 and E2F4 expression was absent in pancreas). As shown, E2F5 and E2F6 were similarly expressed in the WT and DKO pancreas. The last row shows expression of an internal standard (GAPDH) amplified in a reaction parallel to the test genes. (F) Pancreas weight expressed as fraction of total body weight. Shown are means ± SEM for four to six animals per genotype and sex at each time point analyzed. The nonparametric Mann-Whitney U test was used for comparative analysis (*P < 0.05).