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. 2000 Feb;11(2):735–746. doi: 10.1091/mbc.11.2.735

Figure 1.

Figure 1

Association among IRS-1, IRS-2, and Bcl-2. (A) A 180-kDa tyrosine phosphoprotein associated with Bcl-2 in insulin-stimulated IM-9 cells. IM-9 cells stimulated with insulin (100 nM) at 37°C for 5 min were lysed, immunoprecipitated (IP) with C100 anti-Bcl-2 antibody, and immunoblotted (WB) with anti-phosphotyrosine antibody 4G10 (upper panel) or C100 (lower panel). P-Bcl-2, phosphorylated Bcl-2. (B) A 180-kDa tyrosine phosphoprotein associated with Bcl-2 in insulin-stimulated IM-9 cells is IRS-1. IM-9 cells were lysed, immunoprecipitated (IP) with J36 anti-IRS-1 antibody or C100 anti-Bcl-2 antibody, and immunoblotted (WB) with C-20 anti-IRS-1 antibody or C100 anti-Bcl-2 antibody. TCL, total cell lysates. (C) IRS-1 and IRS-2 associate with Bcl-2. Murine IRS-1 or IRS-2 in pRcCMV was cotransfected with human Bcl-2/pUC-CAGGS in 293 cells. Then lysates were immunoprecipitated and immunoblotted with indicated antibodies. +, transfected; −, not transfected. (D) IRS-1 and IRS-2 associate with Bcl-2 in murine hepatocytes. Two milligrams of lysates from murine hepatocytes (C57BL/6J Jcl mice mice) were immunoprecipitated and immunoblotted with the indicated antibodies.