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. Author manuscript; available in PMC: 2010 Oct 1.
Published in final edited form as: Cancer Res. 2009 Sep 29;69(19):7681–7688. doi: 10.1158/0008-5472.CAN-09-2133

Fig 2. ASK1 binds to the N-terminal region of Daxx.

Fig 2

(a) Schematic diagram of deletion mutants of Daxx. Numbers indicate amino acid residues. (b) Mapping of the ASK1-interacting region of Daxx. 293 cells were transfected with the plasmids expressing ASK1-HA and various deletion mutants of FLAG-Daxx, as indicated in (a). Two days post-transfection, cell extracts were subjected to immunoprecipitation with anti-HA antibody, and the precipitated materials were analyzed by western blotting with anti-FLAG antibody. The 20% inputs for immunoprecipitation were analyzed for FLAG-Daxx and ASK1-HA proteins by western blotting as shown (Input WB). (c) ASK1 kinase activity is dispensable for the interaction of ASK1 with Daxx. Cell extracts from 293 cells expressing EGFP-Daxx 70–216 and either ASK1-HA or ASK1 KM-HA were prepared and the expression levels of EGFP-Daxx 70–216 and ASK1-HA were determined by anti-EGFP and -HA antibodies, respectively. Since the expression level of EGFP-Daxx 70–216 was higher in cells when expressed with ASK1-HA than with ASK1 KM-HA, the amounts of lysates for immunoprecipitation were adjusted based on the western blotting data for EGFP-Daxx 70–216 and ASK1. Cell lysates were immunoprecipitated with anti-HA antibody followed by western blotting for EGFP. IP: immunoprecipitation. WB: western blotting.