Expression of Erk1/2 and p-Erk1/2. (A) Effects on immunohistochemistry staining of active form of Erk (p-Erk) by doxorubicin and cisplatin. Multicellular structures of Ishikawa, RL95-2 and KLE cells were cut, fixed and stained with anti-p-Erk antibody. p-Erk was stained green and the nucleus is blue. The scale bar is 50 μm. (B) Western blotting of active form of Erk (p-Erk) and total expression of Erk. 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-Erk) and total expression of Erk from 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).