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. 2005 Jul 18;170(2):295–304. doi: 10.1083/jcb.200409117

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Copyright © 2005, The Rockefeller University Press

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Figure 4. — Expression of active JNK promotes association of Bad with Bcl-2/Bcl-x_L and regulates the phosphorylation state of Bad. (a and b) COS-1 cells were transfected for 18 h with expression vectors for MKK7-JNK and p35 (a) or for Myc–14-3-3ζ (WT or Ser184A) and MKK7-JNK (b) as indicated and were subjected either to immunoblot analysis (IB) with indicated antibodies or to immunoprecipitation (IP) with antibodies to Bad; the resulting precipitates were subjected to immunoblot analysis with indicated antibodies. (b) The asterisk indicates the position of Myc-tagged 14-3-3ζ. (c) COS-1 cells were transfected for the indicated times with expression vectors for MKK7-JNK. (d) COS-1 cells were transfected for 12 h with the indicated plasmids and were incubated for 6 h in the absence or presence of 500 nM okadaic acid (OA). (c and d) Cell lysates were subjected to immunoprecipitation with antibodies to Bad, and the resulting precipitates were subjected to immunoblot analysis with indicated antibodies. (e) COS-1 cells were transfected for 18 h with expression vectors for MKK7-JNK (WT) and 14-3-3ζ Ser184A. Cell lysates were subjected to immunoblot analysis as indicated. The same results were obtained in two independent experiments.