PMC

Mice were fed eRapa (14 ppm) or empty capsules (Eudragit) starting at approximately 8.5 weeks of age until death. (A) Females. Number of Eudragit-fed mice: 8 p53^−/−, 32 p53^+/−, 17 p53^+/+. Number of eRapa-fed mice: 7 p53^−/−, 29 p53^+/−, 21 p53^+/+. (B) Males. Number of Eudragit-fed mice: 15 p53^−/−, 28 p53^+/−, 19 p53^+/+. Number of eRapa-fed mice: 23 p53^−/−, 22 p53^+/−, 21 p53^+/+. (C) Females and males combined. Number of Eudragit-fed mice: 23 p53^−/−, 60 p53^+/−, 36 p53^+/+. Number of eRapa-fed mice: 30 p53^−/−, 51 p53^+/−, 42 p53^+/+.

(A) SAOS2 cells transfected with p53 and increasing concentrations of angiogenin (0.5–5μg Ang-myc plasmid) were Western blotted for phospho p53 (top panel), stripped and reprobed for total p53 (bottom panel). (B) SAOS2 cells were transfected with p53-flag and increasing concentrations of myc-angiogenin and immunoprecipitated with p53 antibody and Western blotted for Mdm2 (top panel), immunoprecipitated with Mdm2 antibody and blotted for p53 (second panel), and immunoprecipitated with Mdm2 and Western blotted with myc (angiogenin; third panel). The lysates were Western blotted for myc (angiogenin) for input control (bottom panel). Actin was used as loading control. (C) SAOS2 cells transfected with p53 and increasing concentrations of angiogenin, or the cells transfected with p53 in the presence of si-angiogenin, were immunoprecipitated for p53 and Western blotted for ubiquitin. Bottom panel shows input p53 blot.

(A) The p53 protein levels in the bone marrow from 129SV^sl p53 P72 and R72 mice were determined by the Western-blot assays. (B) The lifespan of 129SV^sl p53 Hupki mice, including both p53 P72 and p53 R72 mice (median survival: 740 days; n = 381). (C) The lifespan of C57BL/6J Hupki mice, including both p53 P72 and p53 R72 mice (median survival: 490 days; n = 306).

(A) HCT116 cells were left untreated (Control) or treated with AS, or transfected with a control shRNA construct, or AS plus a control shRNA construct, p53 shRNA, or AS plus p53 shRNA. After 24 hours of treatment cell lysates were prepared and probed with antibody against NFATc3, PML, p53 and β-actin. (B) HCT116 cells were left untreated (Control) or treated with AS, or transfected with p53 shRNA, or AS plus p53 shRNA. After 24 hours of treatment cell lysates were prepared and probed with antibody against p53, NFATc4 and β-actin. (C) HCT116 cells were left untreated, or transfected with a empty vector, or with pcDNA3.1-p53 overexpression construct. After 24 hours of treatment cell lysates were prepared and probed with antibody against p53, NFATc3 and β-actin. (D) Same treatment as in C and lysates probed with antibody against p53, NFATc4 and β-actin. (E) RKO and HCT116 cells were untreated or transfected with a NFATc3 shRNA construct and lysates probed with antibody against p53, MDM2, NFATc3 and β-actin. (F) HCT116 cells were untreated or transfected with an empty vector or NFATc3 shRNA construct and lysates probed with antibody against NFATc3, NFATc4, and β-actin. (G) HCT116 cells were left untreated, transfected with an empty vector or NFATc2 shRNA construct, lysates were probed with antibody against NFATc2, p53, NFATc4, and β-actin. (H) HCT116 cells were left untreated, transfected with an empty vector or p53 shRNA construct, lysates were probed with antibody against p53, NFATc1 and β-actin. kDa, molecular weight.