Histology and immunohistochemical staining for EC markers and Akt/4E-BP1 in tumors formed from cell injections. Tumors formed after injection of 1 × 106 cells in the right and left dorsal area of the trunk of 3-week-old male KSN/Slc mice. (A- D) Histological features of formed tumors (A: JuA1, B: JuB2, C: JuB4, D: Re21). Hematoxylin and eosin staining; bars = 50 μm. The neoplastic cells had spindle to polygonal-shaped cytoplasm with oval nuclei, forming some areas of vascular clefts of channels. (E-S) Immunohistochemical results of CD31, vWF, Ki-67, p-Akt Ser473, p-Akt Thr308, and p-4E-BP1 Thr37/46 in the formed tumors. (E) The membrane of the tumor cells show positive staining with CD31 (JuA1). ( F) The cytoplasm of the tumor cells show positive staining with vWF (Re21). ( G) The positive staining of Ki-67 MIB-1 clone in the nuclei of tumor cells indicates that the tumor is not derived from the mice (JuB2). ( H- K) All HSA tumors that developed showed moderate (I: JuB2, J: JuB4, and K: Re21) to strong (H: JuA1) expression for p-Akt Ser473 in the cytoplasm and nuclei. (L- O) HSA tumors that developed showed moderate (N: JuB4) to weak expression (L: JuA1 and M: JuB2), and one cell line (O: Re21) showed no expression of p-Akt Thr308. (P- S) All HSA tumors showed strong cytoplasmic and nuclear expression of p-4E-BP1 Thr37/46 (P: JuA1, Q: JuB2, R: JuB4, and S: Re21). Immunohistochemical staining; bars = 50 μm (G) and 25 μm (E, F, H- S).
Murai et al. BMC Veterinary Research 2012 8:128 doi:10.1186/1746-6148-8-128