Expression of Akt and p-Akt. (A) Effects on immunohistochemistry staining of p-Akt by doxorubicin and cisplatin. Multicellular structures of Ishikawa, RL95-2 and KLE cells were cut, fixed and stained with anti-p-Akt antibody. p-Akt was stained green and the nucleus is blue. The scale bar is 50 μm. (B) Western blotting of active form Akt (p-Akt) and total expression of Akt. After 48 hours treatment with anticancer drugs, cells were lysed and proteins were separated and analysed by SDS-PAGE and Western blotting. The active form (p-Akt) and total expression of Akt of multicellular structures of Ishikawa (B:a-b,), RL95-2 (B:e-f), KLE (B:i-j) and cell monolayers of Ishikawa (B:c-d), RL95-2 (B:g-h), KLE (B:k-l).