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. 2013 Dec 2;4(12):2523–2531. doi: 10.18632/oncotarget.1550

Figure 4. Oncogenic K-Ras dependent p53 endocytosis in vivo.

Figure 4

(A) RP53 was endocytosed into MEF cells obtained from K-Ras transgenic mouse (K-Ras LA2; TgK), but not from wild type mouse. After incubation with RP53 for indicated time, MEF cells were analyzed by western blot. Actin was used as a loading control. (B) Internalization of RP53 was found in K-Ras transformed MEF cells. MEF cells from wild type mouse were transformed with K- and H-Ras, and treated following indicated time. (C) Experimental scheme of in vivo study. Stabilized MEF cells expressing K- and H-Ras were inoculated into 8 week-old wild type C57BL/6 mice through intraperitoneal (i.p.) injection. After 3 weeks, mice were injected with 1mg/ml of His and His-p53 (RP53) 24 h before sacrifice. (D) Tumor mass was obtained from peritoneum of pancreas. Histological analysis was performed by standard hematoxylin and eosin (H&E) staning protocol. (E) Injected RP53 was endocytosed into tumors of K-Ras, but not H-Ras transformed MEF cell. Histological analysis was performed by immunohistochemistry(IHC) using His antibody to verify RP53 endocytosis.