Figure 5. eEF2K Promotes Resistance of Tumors to Caloric Restriction-Induced Cell Death In Vivo.

(A and B) Tumor volumes of NIH 3T3 Ras^V12/MSCV and Ras^V12/eEF2K (A) or EN/MSCV and EN/eEF2K (B) xenografts implanted subcutaneously in nu/nu mice. Mice were fed either AL or placed on CR diets (n = 10 mice/group).

(C) Hematoxylin and eosin staining (H&E) of tumor xenografts from (A). Black arrows indicate regions of necrosis. Results are expressed as percentages of necrotic/total tumor areas (n = 3 mice per group).

(D) Immunohistochemical (IHC) staining of tumor xenografts from (A) with antibodies against cleaved caspase-3. Graphs indicate percent of total cells that are positive for cleaved caspase-3 (n = 3 mice per group).

(E) IHC staining of tumor xenografts from (A) with anti-phospho-eEF2 antibodies.

(F) Immunoblot analysis of tumor lysates of tumor xenografts from (A).

(G) Tumor volumes of eEF2K^+/+ Ras^V12 and eEF2K^−/− Ras^V12 xenografts implanted subcutaneously in nu/nu mice. Mice were fed either AL or placed on CR diets (n = 10 mice per group).

(H) H&E staining of tumor xenografts from (G). Black arrows indicate areas of necrosis. Results are expressed as percentages of necrotic/total tumor areas (n = 3 mice per group).

(I) IHC staining of tumor xenografts from (G) with antibodies to cleaved caspase-3. Graphs indicate the percent of total cells positive for cleaved caspase-3 (n = 3 mice per group).

Where shown, data are reported as means ± SEM with indicated significance (*p < 0.05 and **p < 0.01; ns, nonsignificant). See also Figure S6.