Figure 9. Deficient eIF2αP inhibits growth and promotes senescence of doxorubicin treated human tumors in mice. (A,B) HT1080 WT and KI tumor cells were injected subcutaneously in the flanks of 10 female nude mice for each group. Each mouse received two subcutaneous injections (1×105 cells per injection site) in the abdomen proximal to the rear limbs (n=2×5=10). After injection tumors were left to grow to a measurable size and half of mice (n=5) from each group were treated with placebo and the other half with 4 mg/kg doxorubicin. Tumor growth was monitored for 40 days. Asterisks indicate the time points of doxorubicin injections. (C) At the endpoint of the experiment, tumors were excised from the mice and the mass of each tumor was determined. Histograms represent the average mass of tumors. (D) Equal-sized pieces of tumors were cut from HT1080 WT and KI tumors and subjected to SA-β-Gal staining. (E) Tumor sections from doxorubicin treated WT and KI tumors were subjected to SA β-Gal and H&E staining. (F) The levels of eIF2αP in the WT and KI tumors was assessed by staining of tumor sections with phospho-specific antibodies against Ser51.