Fig. 4. Effect of Mcl-1 overexpression on H<sub>2</sub>O<sub>2</sub>-induced JNK activation and apoptosis. (A) Expression of Mcl-1 protein in several.

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Fig. 4. Effect of Mcl-1 overexpression on H<sub>2</sub>O<sub>2</sub>-induced JNK activation and apoptosis. (A) Expression of Mcl-1 protein in several clonal cell lines by immunoblotting. (B) Sensitivity of Mcl-1 overexpressed cells and vector control cells to H<sub>2</sub>O<sub>2</sub>-induced apoptosis. Briefly, AGS/mcl-1-1, AGS/mcl-1-5, or AGS/neo cells were exposed to 1 mM H<sub>2</sub>O<sub>2</sub> for various periods of time. The apoptotic cells were quantified by the flow cytometric analysis of Annexin-V-stained cell samples. Data are the mean of two highly reproducible experiments. Bar, SD. (C) Mcl-1 overexpression inhibits JNK activity elicited by H<sub>2</sub>O<sub>2</sub>. Mcl-1 transfectants and neo control cells were treated with 0, 1, 2, or 3 mM H<sub>2</sub>O<sub>2</sub> for 2 h, after which cells were harvested and the cytosolic fraction analyzed for JNK1. The kinase activities were determined by immune complex kinase assay and the protein levels were determined by immunoblotting as described in Materials and methods. GST-c-jun was used as a substrate for JNK1. From: IL-6 inhibits apoptosis and retains oxidative DNA lesions in human gastric cancer AGS cells through up-regulation of anti-apoptotic gene mcl-1 Carcinogenesis. 2001;22(12):1947-1953. doi:10.1093/carcin/22.12.1947 Carcinogenesis | © Oxford University Press 1